FISH. FISH was performed as previously described.13 At least 300 cells were counted to determine things the frequency of nuclei having ribonuclear foci. Detection of unpaired single-stranded DNA. Genomic DNA from HT1080 cells were subjected to bisulfite modification for 16 hours at 37 ��C by using EpiTect Bisulfite Modification Kit (Qiagen). Two hundred nanogram of bisulfite-modified DNA was amplified for 36 cycles with the same primers as small-pool PCR, then tailed with 3��-terminal deoxyadenocine by Go Taq polymerase (Promega, Madison, WI). PCR products were electrophoresed on agarose gels, purified, and cloned into pGEM-T Easy vector (Promega). The DNA sequencing was performed with at least 10 individual clones by Big-Dye v.3.1 terminators (Applied Biosystems). SUPPLEMENTARY MATERIAL Figure S1.
Histograms of repeat length distributions in mouse muscle treated with the gapmer LNA-ASO or PBS. Figure S2. Histograms of repeat length distributions in mouse muscle treated with the mixmer LNA-ASO or PBS. Table S1. Repeat instability in individual XXL mice. Acknowledgments This work comes from the University of Rochester Paul D. Wellstone Muscular Dystrophy Cooperative Research Center (NIH/NS048843) with support from the National Institute of Health (AR049077, AR48143); the Saunders Family Fund, the Muscular Dystrophy Association (M.N.); Run America Foundation, and postdoctoral fellowships (to M.N.) from the Cell Science Research Foundation and the Uehara Memorial Foundation. The authors thank Christopher Pearson for critical reading of the manuscript. Supplementary Material Figure S1.
Histograms of repeat length distributions in mouse muscle treated with the gapmer LNA-ASO or PBS. Click here for additional data file.(586K, pdf) Figure S2. Histograms of repeat length distributions in mouse muscle treated with the mixmer LNA-ASO or PBS. Click here for additional data file.(595K, pdf) Table S1. Repeat instability in individual XXL mice. Click here for additional data file.(35K, doc)
Uveal melanoma is the most common primary malignant intraocular tumour in adults. In addition, up to 40% of patients die within 5 years, usually due to hepatic micrometastases.1 2 The most common sites of metastases are the liver (93%), lung (24%) and bone (16%), with 87% of patients with metastasis having multiple sites involved.3 Increased size of tumour has been associated with a worse prognosis. Metastatic spread occurs haematogenously and we have previously reported that murine uveal melanoma cells express and secrete VEGF both locally and at distant sites of metastasis.4 Others have also noted local VEGF secretion within human uveal melanoma.5 VEGF secretion stimulates angiogenesis Entinostat and can enhance metastatic potential.