Figure 5

Figure 5

Telomerase inhibition enhanced the expression and nuclear localization of cyclin-dependent kinase inhibitor 1A (CDKN1A, P21). (A,C) Cells were treated with 30 μM BIBR or DMSO as a control for 24 h. Levels of relative mRNA expression for P21 in 4134/Late (A) and BL41 (C) are shown. Data represent the mean and SD (bar) from three separate experiments. (B,D) 4134/Late (B) and BL41 (D) cells treated with 30 μM BIBR or DMSO as a control for 24 h were processed to obtain cytoplasmic and nuclear extracts. The indicated proteins were probed by Western blotting and representative blots are shown. α-tubulin and TRF2 were used as loading controls for the cytoplasmic and nuclear fractions, respectively. The original Western blots are shown in File S1. Graphs show the values in arbitrary units of densitometric analysis performed with ImageJ software. Data represent the mean and SD (bar) from three separate experiments. (E,F) Representative immunofluorescence images showing P21 (green), PI (nuclear marker, red), and their merge in 4134/Late (E) and BL41 (F) cells treated with 30 μM BIBR or DMSO as a control (magnification 20× unless specified). Graphs show the values in arbitrary units of densitometric analysis performed with ImageJ software. Data represent the mean and SD (bar) from two separated experiments, scale bars: 100 μm in 20× and 5 μm in 63× images. A significant difference between values in BIBR-treated vs. DMSO-treated cells is shown: * p < 0.05; ** p < 0.01; *** p < 0.001; ns: not significant.