For determnatoof doxorubcnduced O2N2 formaton, cells were plated at a densty of 1|106 cells ml and pre ncubated wth 50 mMhydro Cy5 dye resuspended DMSO for 15 mn.Right after pre ncubaton, ten mM doxorubcwas added to respectve wells and knetc fluorescence readngs were takng wth the mcroplate reader each and every ten mfor 1hr.Unstmulated cells, pre ncubated wth and wthouthydro Cy5 dye, and phenol red absolutely free meda, pre ncubated wth selleck and wthouthydro Cy5 dye and doxorubcn, respectvely, had been implemented as controls.All values reported are the average of 3 or a lot more ndependent bologcal replcates 2 traditional error.Statstcal sgnfcance s primarily based upothe crtera of p,0.05 to get a College students check.Fgure S1 Pgactvty the EU1 and EU3 cells are equvalent and nosgnfcant.Dye efflux characterzatofor ALL and AML cell lnes ndcatng that the doxorubcresstant EU1 cells along with the doxorubcsenstve EU3 cells are not sgnfcantly dfferent, regardng ther Pgactvtes, from the PgAML cell lne.Fgure S2 Doxorubctransport for EU1 and EU3 cells are equvalent.
Extracellular doxorubcdepletofor doxo rubcresstant Ostarine EU1 and doxorubcsenstve EU3 cells.Fgure S3 Basal NADlevels are sgnfcantly dffer ent betweethe EU1 and EU3 cells.Relatve basal ntracellular doxorubcresstant EU1 and doxo rubcsenstve EU3 cells determned by absorbance readngs.Fgure S4 Senstvty analyss of model parameters and speces concentratons.Chosen parameters and speces ntal condtons have been systematcally perturbed and the model predcted results of those varatons oqunone doxorubcaccumulaton, NADdepleton, and superoxde productowere assessed.The ntal values applied for the senstvty analyss, had been takefrom the EU1 Res cell model with the 10 mM doxorubcconcentratocondton.These values had been thencreased by 10% or decreased by 10%, ndependently, and themodel smulatons had been carred out ndcates the parameters for whch the knetc fee constants had been vared and ndcates the parameters for whch the ntal concentratons have been vared.Model senstvty analyss was performed for any ten mM doxorubctreatment regmen.
Normalzed senstvty coeffcents have been calculated to quanttatvely characterze the effect of each parameter perturbatooqunone doxorubcaccumulaton, NADdepleton, and superoxde producton, respectvely.The

ordinary zed senstvty coeffcents are showFgure S4. Two dmensonal monolayer cell cultures representhghly reductonst versions of epthelal cells and epthelal cancers, due to the reduction of physologcal extracellular matrx oartfcal plastc surfaces, andhgh serum concentratons.Consequently, cells drop appropriate propertes, such as dfferentaton, polarzaton, cell cell communcatoand extracellular matrx contacts, whe woundhealng, nflammatory processes, andhyper prolferatoare artfcally promoted.monolayer culture of prostate cancer lnes, thehomeostass of undfferentated tumor stem cells by way of basal, transt amplfyng and termnally dfferentated,hormone senstve lumnal cells depends ocell culture condtons, calcum and serum concentraton, and only poorly represents tumor cell bology vvo.