BRG1 does not alter p16INK4a driven senescence. WMM1175_p16INK4a cells, BRG1 silenced (clone X1, left panel) or NS (clone E1, right panel), were exposed to 4 mM IPTG over a five-day period and analyzed by FACS analysis, Western blot and imunocytostaining: A 50 μg of total cell lysate were immunoblotted and probed for p16INK4a, phospho-pRb (pRbSer807/811) and as a loading control β-actin. B The accumulation of p16INK4a, the cell proliferation marker Ki67, chromatin condensation (DAPI) and the appearance of SA-β-gal was analyzed by immunocytostaining in WMM1175_p16INK4a. Enlarged images of cells (indicated with arrows) show DAPI-stained chromatin foci. Histograms correspond to the average ± s.d of at least two independent induction experiments from a total of at least 500 cells. LM, light microscopy. C FACS analysis by Forward Scatter (FSC) and Side Scatter (SSC) of clones demonstrate the senescence associated increase of cell size (FSC) and granularity (SSC) upon p16INK4a induction.