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. 2015 Aug 11;6(29):27130–27145. doi: 10.18632/oncotarget.4825

Figure 4. The synergistic regulation of hTERT promoter activity, hTERT expression, telomerase activity and cell proliferation in H1299 cells with overexpression of RFPL3 and low expression of CBP.

Figure 4

A. Streptavidin-agarose bead pulldown assay with hTERT promoter (−378 to +60) as probes was done in lung cancer cells (H1299 cells stably expressing RFPL3) treated by non-specifc siRNA or CBP specifc siRNA or CBP-specifc inhibitor(C646). The level of RFPL3 in the pulled down proteins was determined by immunoblot (NSP: non-specifc control siRNA). B. H1299 cells with stable overexpression of RFPL3 were co-transfected with CBP-specific siRNA1, or CBP-specific siRNA2 or CBP inhibitor and the plasmids of hTERT promoter driven-luciferase for 48 hours followed by luciferace assays. The relative luciferase intensity per μg protein was calculated in the treated cells. C. H1299 cells with stable overexpression of RFPL3 were transfected with CBP siRNA or incubated with CBP-specific inhibitor for 48 hours, and the expressions of hTERT, RFPL3 and CBP proteins in the nuclear extracts of these cells were examined by western blot. D. The telomerase activity in the cells treated as above was measured. E. Cell viability of these cells treated as above was measured by MTT assay. All of the measurements represent the means ± SE of three independent experiments. *P < 0.05, **P < 0.01, significant differences between treatment groups and control groups.