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. 2014 Sep 11;136(7):1546–1558. doi: 10.1002/ijc.29171

Figure 2.

Figure 2

Absence of hTERT expression in TG20 ALT cells. (a) hTERT mRNA expression in telomerase-positive GSCs (TG16, TG1N, OB1 and TG10) and ALT cells (TG20 and SAOS-2). Values are presented as the mean ± SEM of two independent experiments performed in duplicate. (b) The same cell lines as in (a) were evaluated but for hTERC mRNA expression. (c) Representative methylation-specific PCR (MSP) results for the proximal region and WT1 binding site of the TERT promoter in SAOS-2 and TG20 ALT cells and in TG1N and TG16 telomerase-positive cells. Bands in the M and U lanes represent methylated and unmethylated PCR products, respectively. (d) mRNA expression of hTERT, TES, and RUNX3 in TG20 cells after treatment with 2.5 or 5 µM 5-azacytidine (5-aza), relative to their expression in control DMSO-treated cells. (e) WT1 protein levels determined by Western blot analysis for the indicated cell lines. β-Actin was used as a loading control. (f) Comparison of expression ratios of WT1 protein vs. β-actin in the different cell lines. Experiment was repeated twice using samples from independent cell cultures.