Figure 5.

The Rb/E2F pathway mediates plasma membrane depolarization‐induced senescence. (a) The transcription Factor ChIP‐seq Uniform Peaks database from the Encode consortium was explored for seven ChIP‐seq experiments performed on E2F1 or E2F4 transcription factors. We counted the number of experiments showing a ChIP‐seq peak with a minimal score of 500 in the promoter of the top 30 genes identified in transcriptome analysis in four different classes: downregulated mitotic genes (Dw‐GO mitosis); downregulated genes without GO enrichment (Dw); upregulated genes (Up); no downregulated mitotic genes (GO mitosis no Dw). (b,c) HEC‐TM cells were infected with a control (ctrl) or E7 encoding retroviral vector (E7) and selected using neomycin. (b) Cells were then treated or not with 65 mM KCl (KCL) for 8 or 24 hr. RNAs were prepared, RT‐qPCR was performed on the indicated mitotic genes, and the results were normalized against GAPDH mRNA levels. (c) The day after seeding, cells were treated daily with 65 mM KCl. After 8 d, cells were fixed and stained using crystal violet. Experiments were performed at least twice. Statistical analysis was performed with the Student's t test, ** means p < .01, ***p < .001