Figure 5
From: The chromatin remodelling factor BRG1 is a novel binding partner of the tumor suppressor p16INK4a
BRG1 does not alter p16INK4a cell cycle regulation. A 50 μg of nuclear lysates from WMM1175_p16INK4a clones with stably integrated siRNA targeting BRG1 or a non-specific (NS) control siRNA were probed for BRG1 and topoisomerase II (Topo II) as a loading control. B 50 μg of total cell lysates extracted from WMM1175_p16INK4a cells stably expressing either a BRG1-specific siRNA or a non-specific (NS) siRNA molecule, as indicated, were treated with PBS (-) or IPTG (+) for 24 h and probed for p16INK4a and β-actin. C Cell proliferation was determined by MTS assay. D A proportion of the IPTG/mock treated cells were analyzed for changes in cell cycle distribution. Percent S-phase change was calculated (percent S-phase mock treated cells – percent S-phase IPTG treated cells) × 100/percent S-phase mock treated cells. E The same clones were seeded at low density (103 cells/7.5 cm plate) and p16INK4a expression was induced with 4 mM IPTG or cells mock treated and colony forming ability was assayed after 14 days.