Further, Mino treatment method considerably reversed the perturb

Even more, Mino therapy drastically reversed the perturb ation of ZO 1 localization, as indicated by masked image scoring carried out by impartial evaluators. Minocycline therapy inhibited expression of ischemia reperfusion responsive genes connected with neuroinflammation, but not individuals linked with astrogliosis following ischemia reperfusion To examine the result of Mino therapy to the inflam matory response following IR we utilized qRT PCR analysis to examine the expression amounts of 25 mRNAs at 48 h following IR in rats systemically treated with or with no Mino as described over. The mRNAs examined were picked from a previously obtained transcriptomics data set for this rat IR model, with preference for genes connected with irritation and astrogliosis.

For comparison, the induc tion of expression of each of these mRNAs in retinas four h following intravitreal injection of 1 ug eye of lipopoly saccharide was also examined. Retinal expression of 21 of these mRNAs had been significantly altered by IR, with twenty order inhibitor mRNAs greater from 89% to 47. six fold and only glutamate ammonia ligase appreciably decreased 55% by IR. Significantly improved mRNA expression was observed for many professional inflammatory genes, which includes, C X C motif chemokine ligand motif chemokine ligand. In contrast, expression of 3 mRNAs which might be characteristic of classical inflamma tion, C X C motif chemokine ligand ten, inducible nitric oxide synthase two and prostaglandin endoperoxide synthase 2 cyclooxygenase two, were not drastically altered at 48 h right after IR.

Nevertheless, the retinal amounts the original source of those three mRNAs have been appreciably increased by LPS injection by 4700 fold, 63 fold and 9. 4 fold, respectively. Expression of mRNAs representing two markers of astrogliosis, glial fibrillary acidic protein and vimentin, have been significantly improved by six. 8 fold and 3. 0 fold, respectively, by IR. Neither of these mRNAs was responsive to LPS injection. Evaluating mRNA ranges in sham taken care of eyes from rats handled with and devoid of Mino exposed that drug treatment method alone appreciably impacted the expression of five mRNAs, together with IL6, C C motif chemokine ligand 5, endothelin 2, CCL3 and also the lectin galectin binding soluble 3 see. Mino also considerably inhibited the IR induced expression of 5 genes. These incorporated ICAM 1, lipocalin 2, serpin peptidase inhibitor clade A member 3 N, TNF receptor superfamily member 12A and CCL2. The inhibitory ef fects of Mino therapy over the IR responses of various other pro inflammatory genes had been nearly substantial. These integrated, CXCL2, IL6, IL1B, and TNF. In contrast, Mino remedy did not significantly affect the IR responses of GFAP and VIM, with calcu lated inhibitions of 10% and 2%, respectively.

These effects are analogous to these obtained in HeLa cells handl

These results are analogous to these obtained in HeLa cells handled with all the pan caspase inhibitor, ZVAD. We con clude that Bcl 2 over expression renders HeLa cells resistant to MiTMAB induced cell death, but to not MiTMAB induced cytokinesis failure. The involvement of caspase 9 and Bcl 2 more indicate activation from the intrinsic apoptotic pathway. MiTMABs induced cell death takes place by way of the intrinsic apoptotic pathway The activation of a further initiator caspase, caspase eight, was also detected in cells handled with MiTMABs. In contrast to cas pase 9, caspase eight is actually a component on the extrinsic apopto tic pathway and it is hence typically activated following stimulation of cell surface receptors. Once activated, it cleaves the pro apoptotic Bcl two member of the family, Bid, which in turn stimulates the intrinsic apoptotic pathway to advertise cytochrome c release from mitochondria.

However, caspase eight can also be activated by cas pase 9 3 within a feedback loop to amplify the currently active intrinsic pathway. For that reason, we sought to find out if activation of caspase inhibitor Wnt-C59 8 in response to MiTMABs takes place following stimulation of your extrinsic pathway and or via intrinsic cell death signals. We to start with investigated the ability of MiT MABs to induce apoptosis inside the presence on the cas pase eight selective inhibitor IETD. Should the intrinsic pathway was solely induced by caspase 8, inhibiting caspase eight alone ought to block cytochrome c release and subsequent cell death. On the other hand, inhibition of caspase 8 only blocked apoptosis by about 40%, in striking contrast to your effect in the pan caspase inhibitor, ZVAD.

IETD treatment also resulted in only a modest raise in polyploid cells, presumably mainly because a substantial proportion of cells that failed cytokinesis had been ready to undergo apopto sis. These findings recommend that activation of caspase 8 induced by MiTMABs is by way of the intrinsic pathway. Bcl 2 above expression blocks cell death upstream of caspase 9 selleck chemicals and 3 activation and as a result caspase eight cleavage really should be prevented in HeLa Bcl two cells if it is activated solely via the intrinsic pathway. In line with this particular plan, we didn’t detect cleaved caspase eight in MiTMAB taken care of HeLa Bcl two cells. In contrast, caspase eight cleavage was detected in both HeLa and HeLa Bcl two cells exposed to UV, a acknowledged stimulant in the extrinsic pathway. We conclude that MiTMABs induce apoptosis via the intrinsic apoptotic pathway and this consists of activation of caspase 8 by way of a suggestions amplification loop. The apoptotic response of cancer cells to MiTMABs appears to correlate with expression of Bcl two and Mcl 1 anti apoptotic proteins We up coming aimed to verify if MiTMABs induce apoptosis in other cancer cell lines.

On its recruitment to the cellular membrane by means of receptor

On its recruitment to the cellular membrane through receptor mediated activation, the p110a catalytic sub unit of PI3K phosphorylates phosphatidylinositol 4,five bisphosphate in the 3 place in the inositol ring, creating PIP3. PIP3 recruits phospholipid binding domain containing proteins, specifically AKT, to your plasma membrane. BGB324 Phosphorylated AKT, the main downstream effector of PI3K signaling, moves in the cytoplasm on the nucleus to initiate its downstream effects. This cascade, including activa tion of your mammalian target of rapamycin and its downstream effectors, p70S6 kinase and 4E binding protein 1, impacts a variety of cellular pro cesses, together with proliferation and BGB324 motility, which clinically translate into endocrine and chemotherapy resistance and worse cancer specific survival.

The PI3K AKT pathway is negatively regulated by PTEN, a lipid phosphatase that removes the three phosphate from PI P2 and PI P3, hence inactivating the signaling cascade. Therefore, loss of PTEN contributes on the purchase Wortmannin activation of the PI3K AKT signaling cascade via inhibition of degradation of each PI P2 and PI P3. To date, alterations and activation of the PI3K AKT pathway BKM120 are properly established during the initiation and professional gression of extracranial human BC. How ever, the contribution of this vital signaling pathway to the pathogenesis of BCBMs has but for being thoroughly elucidated. This really is of clinical value as modest molecule inhibitors of your PI3K AKT mTOR pathway are in growth and demonstrate promising exercise within the remedy of major brain tumors, suggesting sufficient blood brain barrier penetration to elicit therapeutic results.

On this study, we quantitated the expres sion of BKM120 the PI3K pathway biomarkers p AKT, p S6, and PTEN, and evaluated the prognostic implications, pri marily total survival and survival just after BCBMs, of PI3K activation status in BCBMs. As secondary, exploratory end factors, we evaluated the associations between PI3K pathway activation and time to distant recurrence and time to BCBM. Finally, similar analyses had been also carried out amongst the subset of sufferers with triple damaging BCBM. Elements and techniques Sufferers BCBMs, together with a subset with matched pri mary BCs, from 52 sufferers treated with the Uni versity of North Carolina at Chapel Hill and Duke University selleckchem in between 1991 and 2008, were studied. Clinical data, such as age, race, stage of main BC at diagnosis, therapy historical past, recurrence, and vital status were obtainable for 50 sufferers. Provided the ret rospective nature of clinical data collection, complete information was not offered for all 50 sufferers, hence, denominators may fluctuate throughout the posting. This study was accredited, and waivers of consent have been granted by Institutional Review Boards at each UNC and Duke.

In studies from our laboratory employing a LCC6 cell line complet

In research from our laboratory utilizing a LCC6 cell line permanently transfected using the Her2 gene, we now have been in a position to demon strate the Her2 optimistic variant exhibit enhanced survival beneath strain, overproduction of VEGF, activation of nuclear issue B and in vivo sensitivity to Tz, success which are constant with precisely what is acknowledged about Her2 favourable breast cancer models. Therefore, we feel it can be justifiable to utilize these cells like a model breast cancer cell line, notably when the success obtained making use of this cell line are confirmed with other breast cancer cell lines. LCC6, LCC6Her2, LCC6luc, KPL 4, BT 474, MDA MB 468, MCF 7 and MCF 7Her2 cells had been maintained in Dulbeccos modified eagles medium higher glucose supple mented with L glutamine 5 mM penicillin streptomycin, and 10% FBS.

SKBR3 cells were maintained in McCoys 5a medium supple mented with L glutamine, 5 mM penicillin streptomycin, and 10% FBS. All cells have been maintained at 37 C and 5% carbon dioxide inside a humidified atmosphere. Cell viability assays Metabolic action of breast cancer cell lines incubated inside the presence of different therapeutic agents was determined selleck chemical working with Alamar Blue assays according to your suppliers tips. Briefly, 6000 cells nicely seeded in triplicate onto 96 properly flat bottom tissue culture plates had been allowed to adhere on the substratum for 24 hrs below typical growth circumstances. Serial dilutions of personal medicines, 267 drug combinations and vehicle controls diluted in acceptable cell culture medium were then extra to the wells and cells had been grown for an addi tional 72 hrs.

To assess cell viability, cells had been then incu bated with 10% resazurin remedy for four hrs at 37 C and fluorescence was measured at 560 590 nm employing an Optima fluorescence plate reader. Relative fluorescence determined from drug taken care of cells was normalized to fluorescence established from control cells and information is shown as over here percentage relative cell viability com pared with automobile handled management cells. Background fluorescence was subtracted from all samples and success of experiments conducted in journey licate are indicated. Drug combination results median effect principle To find out irrespective of whether numerous 267 drug combinations had resulted in synergistic, antagonist, or additive effects, the median effect principle approach to Chou and Talalay was applied to find out blend index values.

Within this respect, modification of NF B levels may perhaps modi

Within this respect, modification of NF B amounts may modify the concentration of the variety of apoptotic connected aspects. Our final results recommend that caffeic acid might bind towards the AhR, getting an inhibitor of its action, consequently decreasing the transcription and activity of CYP1A1, each in basal and TCDD handled cells. This implies either a direct effect to the enzyme molecule or perhaps a competitors for that AhR with all the endogenous ligand from the AhR. This latter hypothesis looks far more probable as, in our experimental situations, precisely the same inhibitory pattern was observed in either case. To our knowl edge, this really is the very first report indicating an interaction of phenolic acids together with the AhR. It had been lately proven that the result of TCDD is exerted via binding to AhR.

AhR TCDD complicated in flip induces CYP1A1, leading to a significant boost from the DNA binding action of NF B and apolipoprotein 1, and selleck chemical CX-4945 a sustained activation of those two transcription things. It is of note that this activation was blocked by antioxidants. On the contrary, activation from the Fas receptor induces the phosphorylation of NF B transcription element, leading to induction of apoptosis in the number of numerous cell sorts. Contemplating the purpose of NF B in cancer cell apoptosis, it is tentative to hypothesize that caffeic acid may well act by inhibiting this pathway. This hypothesis is even further supported by the stimulation impact of caffeic acid on pro apoptotic Fas receptor. In an hard work to uncover other pathways of apoptosis, concerned from the professional apoptotic actions of phenolic acids on T47D cells, we have now also examined their effects about the members of the other key family members of apoptosis relevant components, the Bcl two proteins.

Bcl two proteins are strongly expressed in human breast cancer cells, including the T47D cells. selelck kinase inhibitor Remarkably, both phenolic acids ele vated the protein written content of your apoptosis stopping Bcl 2 protein. It is actually doable that a Bcl two related mechanism is activated to quick phrase counteract the pressure signals gen erated by the apoptosis inducing component FasL to be able to rescue the cells from programmed death. Yet another possi bility is Bcl 2 relevant anti apoptotic proteins, in the outer mitochondrial membrane, improved to counteract the pro oxidant results of phenolic acids locally. Conclusions The existing operate suggests that phenolic acids exert a direct antiproliferative action. This action is evident at low concentrations, comparable with people identified in biological fluids immediately after ingestion of foods rich in phenolic acids. Fur thermore, the direct interaction using the AhR, the interaction with all the NOS technique as well as pro apoptotic effect of phenolic acids give insights about their mode of action.

The PCR primers and conditions made use of for mouse Col2a1, Mmp3

The PCR primers and ailments applied for mouse Col2a1, Mmp3, Mmp13, Ptgs2, Nos2 and Gapdh have been previously described. The PCR primers for Lrp5 and Lrp6 have been as follows, mouse Lrp5, sense, Quantitative RT PCR was performed employing an iCycler and SYBR Premix Ex Taq. Western blot evaluation Complete cell lysates had been prepared with lysis buffer containing 150 mM NaCl, 1% Nonidet P 40, 50 mM Tris, 0. 2% SDS, 5 mM NaF, a protease inhibitor cocktail in addition to a phosphatase inhibitor cocktail. Proteins were resolved by SDS Page, transferred to nitrocellulose membranes, de tected by incubation using the proper principal antibody as well as a peroxidase conjugated secondary antibody and visualized applying an enhanced chemiluminescence method.

The primary antibodies applied had been bought from ABGENT, EMD Millipore, BD Biosciences, 610408, B catenin, 610154 selleck chemical Santa Cruz Biotechnology and Cell Signaling Technological innovation, 9252, and phosphorylated JNK, 9255, Danvers, MA, USA. Transfection and reporter gene assay Mouse articular chondrocytes had been cultured for 3 days, transfected for four hrs with Lrp5 tiny interfering RNA or pSPORT6 Lrp5 employing Lipofectamine 2000 reagent, then treated with IL 1B, Wnt3a or Wnt7a. A nonsilencing control siRNA and empty vector had been applied as the damaging controls. To deter mine the transcriptional exercise of B catenin Tcf Lef, we utilised a reporter gene assay. Chondrocytes were transfected with 1 ug of reporter gene or management gene and 1 ug of pCMV B galactosidase employing Lipofectamine 2000. The transfected cells had been taken care of with IL 1B, Wnt3a or Wnt7a for 24 hrs, then luciferase acti vity was measured and normalized with respect to transfec tion efficiency.

Statistical examination The nonparametric Mann Whitney U check was made use of to analyze information dependant on ordinal grading systems, including Global Cartilage Fix Society and Mankin scores. For qRT PCR success and apoptotic selleck inhibitor cell numbers, the data had been very first examined for conformation to a normal distribution employing the Shapiro Wilk check, then analyzed by Students t test or analysis of variance with submit hoc exams as ap propriate. Significance was accepted on the 0. 05 degree of probability. Effects Lrp5 is upregulated by way of JNK and NF κB pathways for the duration of IL 1B mediated pathogenesis of chondrocytes We 1st examined the expression amounts of Lrp5 and Lrp6 in the course of the chondrogenic differentiation of mesen chymal cells obtained from mouse embryonic limb buds and subjected to micromass culture.

Equal quantities of total RNA from theront and trophont phases we

Equal quantities of total RNA from theront and trophont stages were pooled. PolyA RNA was chosen and normalized by Evrogen, Inc. The normalized cDNA popula tion was sequenced making use of the Illumina platform, gener ating a hundred bp paired end reads. A complete of 1. 65 × 107 excellent reads were obtained, for a complete of one. 67 Gb of raw RNA seq information. These reads have been aligned to your genome sequence and assembled employing the TopHat suite. Alignments had been even more refined utilizing PASA. Of 24,264 assemblies input into PASA, 24,078 pro duced valid alignments and 23,585 subclusters. Also, 32,606 Sanger ESTs recognized as remaining derived from Ich have been downloaded from NCBI and aligned towards the genome making use of PASA. Of these, 22,483 produced legitimate alignments. Quite a few of the non aligned ESTs matched genes of fish or bacterial origin, suggesting that they’re contaminants.

Assembly with the valid ESTs created four,751 subclusters. Protein coding gene getting To train gene obtaining algorithms, a set of 1,044 gene structures was modeled manually applying the Sanger and Illumina EST alignments and homology to predicted genes of other species, specifically other ciliates. This set was utilized to train 3 ab initio gene prediction professional grams, selleck chemical Augustus, GeneZilla and GlimmerHMM. An original complete set of gene predictions was gener ated determined by the three ab initio algorithms, Ich ESTs, and protein homologies to T. thermophila, P. tetraure lia, Oxytricha trifallax plus a J Craig Venter Insti tute non redundant protein database, aligned applying the AAT and GeneWise packages. Pfam domains were also searched against the genomic sequence.

Evidence through the gene finders, protein and domain homology searches and ESTs had been utilized to refine gene models making use of EvidenceModeler. Superior quality EST alignments selleckchem have been employed to automatically update gene construction annotations working with PASA. Immediately after in depth manual annotation of picked genes, a total of 8,096 gene designs had been created. Automated functional annotation Gene names were computationally assigned by searching protein databases, such as the J Craig Venter Institute Panda comparative database, Panther, Pfam and Uniprot, working with BlastP. A subset with the success was manually reviewed to find out cutoffs that professional duced acceptable names from each of the databases. A subset of gene models was analyzed for correctness and sensitivity to functional assignments. Paralogous households have been computed based mostly upon shared domain composition. A minimum of three paralogs had been required to designate a family. Multivariate evaluation of codon usage was carried out utilizing the codonW bundle as pre viously described. Non coding RNAs Transfer RNAs have been detected utilizing tRNAscan SE with default parameters.

The expression level of transgenic mature ADAM10 is 30% over endo

The expression amount of transgenic mature ADAM10 is 30% above endog enous ranges and in dnADAM10 mice the expression in the catalytically inactive ADAM10 mutant is sevenfold above endogenous ADAM10. ADAM10 activity was deter mined in past studies by quantitation from the APP cleavage item APPs. In ADAM10 overexpressing mice the catalytic activity of ADAM10 towards its substrate APP was greater to about 250%. In mice overex pressing dnADAM10, the endogenous APP cleavage exercise was diminished to about 25% as in contrast to APP mice. For your 1st experimental series with the present examine, female ADAM10, dnADAM10 and FVB N wild variety mice have been investigated, for that 2nd series, female and male ADAM10 APP, dnADAM10 APP and APP mice have been compared.

In each and every case, brains of three five months outdated animals of each group have been dissected and stored in RNA later on at 80 C to prevent RNA degradation. RNA preparation and microarray analyses Complete RNA from full mouse brains was isolated by utilizing the RNeasy Kit, which includes on column DNase selleck chemical I digestion in accordance to the manufac turers recommendations. The high-quality of isolated RNA was controlled through the Lab on Chip Program Bioanalyser 2100. The expression profiling examination for mono transgenic mice was carried out at RZPD.Samples from double transgenic mice were analyzed on the Microarray Facility. In all scenarios, the Mouse Genome 430 two. 0 Array containing 45000 probe sets of 34000 genes was utilized for mRNA expression profiling. Statistical examination and gene annotations For your to start with series 9 gene chip arrays and for the second series 18 gene chip arrays were analyzed.

Data mining was per formed through the use of the ChipInspector analysis software program, which identifies selleck chemicals signifi cant modifications primarily based on single probes. The corresponding transcripts have been then assigned immediately after a user defined quantity of sizeable probes. For all analyses, a transcript coverage greater than three probes was selected. By this method, annotation errors and mistakes brought about through the existence of alternative transcripts are lowered. Just after total intensity normalization of each array, signifi cantly transformed genes have been determined by significance examination of microarrays, using the exhaustive comparison mode at a false discovery fee of 0. 0% for double transgenic, and 0. 5% for mono transgenic mice. For separate analysis of samples from double trans genic female and male mice, a FDR of one. 3% was selected. The resulting gene lists had been restricted to your 600 most strongly regulated genes. Regulated genes were then analyzed with the Bibliosphere program and mapped to Gene Ontology trees in order to determine their biological function. For identification of over represented GO terms, the Bibliosphere software package cal culates a z score for every term.

Visualization was accomplished utilizing the Alexa Fluor 546 con

Visualization was accomplished using the Alexa Fluor 546 con jugated secondary antibody along with a fluorescence mi croscope. Below our culture situations, far more than 99% cells have been good for GFAP in astrocytic culture. Calcium spectrofluorometry A earlier process established for measurement of intracellular Ca2 was modified and followed. In short, 2 five × 105 of astrocytes plated on 22 mm coverslips had been incubated with all the fluorescent Ca2 indicator Fura 2 AM plus pluronic acid in regular physiological saline resolution for twenty min at 37 C. PSS contained, NaCl, KCl, MgCl2, HEPES, D glucose and CaCl2, pH of seven. 4. In some experiments, Ca2 free PSS was used, this solution had the same com place as PSS except that 1 mM of EGTA was added as opposed to CaCl2. All reagents utilized in this assay have been obtained from Sigma Aldrich.

Following a 20 minute wash in dye free PSS at 37 C, coverslips have been placed on the selleck EPZ-5676 stage of an inverted microscope equip ped by using a 40× goal. Cells were exposed to alternating wavelengths of 340 nm and 380 nm for excitation at 6 second intervals. Emission light was passed via a 510 nm filter. An im aging system was utilized to record fluorescence ratios utilizing a CCD camera. The bath chamber was developed to maintain a frequent bath volume and conventional saline PSS was employed to rinse the bath quickly just before experiments. The bath solu tion was static using the exception of alterations in remedy, utilized within 60 s right after PSS rinse, and associated with the addition or removal of agonists and antagonists.

Responses to purinergic application are presented as fluo rescence intensity ratios at excitation wavelengths of 340 to 380 nm versus i was reading this time with all experiments performed at space temperature. Amplitudes of all re sponses in this examine are described as ratiometric values derived in the ratio of excitation wavelengths. ATP induced responses exhibited rapidly and slow com ponents of decay. The time program with the quick initial decay was measured at a point at half amplitude of peak response. The time program with the secondary slower phase of decay was measured at half amplitude of this compo nent. The height of your prolonged phase was determined since the stage of intersection with the element with time at peak response. ATP response in Ca2 cost-free PSS or in conventional Ca2 solution containing Gd3 showed single phase decays from a peak worth with time programs deter mined at half amplitude values of peak. BzATP induced response consisted of a single phase of a slowly create ing improve to a peak level with amplitude of fluorescent ratio utilized being a measure of response.

On the other hand, AKT1 mutation and expres sion status as well a

Even so, AKT1 mutation and expres sion status as well as expression adjustments in other genes on the PI3K AKT pathway did not display any statistically significant association potentially because of the smaller variety of AKT1 mutated instances. mRNA expression amounts of other genes involved during the PI3K AKT pathway were also evaluated, i. e. EGFR, PDK1, PTEN, AKT2 and three, GOLPH3, P70S6K, and WEE1. Markedly high expression that might be caused by gene amplification was observed only in minimal frequency of tumors as exhibits the final colon within the Table one. PTEN underexpression was substantially mutu ally unique with PIK3CA, PIK3R1 and AKT1 muta tions, since it was observed in only one AKT1 mutated tumor and 14 PIK3CA mutated tumors. Ex pression ranges have been also in contrast from the four breast cancer subgroups as shown in Table two.

Interestingly, gene expressions have been deregulated in different strategies from the four subgroups. EGFR underexpression was demon strated in all subgroups, as previously published. P70S6K and selleck chemical AKT1 was predominantly overexpressed in ERBB2 tumors. This enhanced expression of these two genes may very well be linked for the PI3K AKT pathway activated by ERBB2 overexpression. Alternatively, expression alterations in HR ERBB2 tumors might indicate downstream activation from the pathway occurring in spite of the nega tivity of ERBB2. The 4 molecular subgroups of breast cancer therefore appeared to undergo distinct adjustments on the ranges of mRNA expression in the genes in volved during the PI3K AKT pathway. These data would benefit from confirmation at protein degree.

The following stage of examination targeted on selleckchem PI3K constitu ents, particularly PIK3R1 expression and PIK3CA muta tions in relation to expression amounts on the other genes evaluated. Tumors characterized by PIK3R1 underexpres sion were connected with deregulation of other genes involved in the PI3K AKT pathway. PIK3R1 underexpression was negatively connected with PIK3CA mutations and these two parameters have been therefore predominantly mutually unique. In contrast to PIK3R1, deregulation of your expression of genes concerned during the PI3K AKT pathway was virtually solely associ ated with PIK3CA wild sort tumors. Immunohistochemistry Alteration of p85 and PTEN ex pression was also verified on the protein level by im munohistochemistry in randomly picked samples with minimal and substantial mRNA expression. In each scenarios, sam ples exhibiting decreased mRNA expression also presented very low immunohistochemical staining inten sity. Similarly, samples showing typical mRNA expression presented sturdy immunohistochemical staining intensity. The sole exceptions have been two samples stained for PTEN. A good match was as a result obtained among mRNA and protein expression standing for the two PIK3R1 and PTEN.