Figure 5

MUC1 knockdown induces changes in telomere maintenance mechanism. (A) qRT-PCR results showing significantly lower TERT mRNA expression in MUC1 knockdown cells. Data representative results from at least 3 independent experiments and were plotted with average of 3 independent experiments. Error bars, SEM. Student’s t-test. *P < 0.05, **P < 0.01, ***P < 0.001. (B) MUC1 knockdown cells showing significantly reduce telomerase activity. Telomerase activities were analyzed using telomerase PCR ELISA kit and was quantified as ΔA (A450-A690) and then calculated as RTA (%). Data representative results from at least 3 independent experiments and were plotted with average of 3 independent experiments. Error bars, SEM. Student’s t-test. *P < 0.05, **P < 0.01, ***P < 0.001. (C) Blot showing positive formation of C-circles in MUC1 knockdown glioblastoma (GBM) cells. Genomic DNA isolated from control (CTR) and MUC1 knockdown T98G and U373 cells used for the C-circle assay. U2OS and WI-38 VA13 cells were used as the positive control and 293 T cell was used as the negative control. The blots were cropped and full length blots are presented in Supplementary Fig. S3. (D) Telomere restriction fragment (TRF) analysis using T98G and U373 cells infected with shLuc or shMUC1 lentivirus infection. Mean telomere restriction fragments (TRF) detected by southern blot analysis. Data are representative results from at least 3 independent experiments.