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. 2017 Apr 27;8:15034. doi: 10.1038/ncomms15034

Figure 6. ZNF148 depletion results in decreased TERT expression and telomerase activity.

Figure 6

(a) An siRNA directed against the ZNF148 transcript was transfected into A549 (lung, left) and MIA PaCa-2 (pancreas, right) cell lines, and expression of ZNF148, TERT, CLPTM1L, ACTB and GAPDH were assayed by quantitative PCR. Depletion of ZNF148 resulted in consistent reduction of TERT but not CLPTM1L or control gene expression. Expression values were normalized to those from cells transfected with a scrambled control siRNA. Experiments were conducted in triplicate and repeated three times. Mean measures for three independent experiments are plotted; error bars represent s.e.m. (b) siRNAs targeting ZNF148, TERT, the regulatory region encompassing rs36115365 (siRNA3), or a scrambled siRNA control were transfected into A549 (lung, left) and MIA PaCa-2 (pancreas, right) cell lines, and telomerase activity was measured via a telomeric repeat amplification protocol (TRAP). Negative control represents the TRAP assay performed with no cell extracts added. The internal control represents the 36 bp internal standard. (c) siRNAs targeting ZNF148, TERT, or the regulatory region encompassing rs36115365 (siRNA3) were transfected into A549 (lung, left) and MIA PaCa-2 (pancreas, right) cell lines repeatedly (once every four days), and telomere length was measured after 20 days using quantitative PCR for telomere repeat copy number. Data are normalized to those from a scrambled siRNA control. Experiments were conducted in triplicate and repeated three times. Mean measures for three independent experiments are plotted; error bars represent s.e.m.