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. 2016 Oct 1;30(19):2187–2198. doi: 10.1101/gad.286112.116

Figure 4.

Figure 4.

Ectopic expression of ARID1B induces senescence. (A,D,E) IMR90 cells were transduced with either an empty vector, ARID1B cDNA, or RasG12V cDNA and then analyzed for cell growth (crystal violet), SA-β-Gal activity, and BrdU incorporation (A) as well as IF staining (D) using the indicated antibodies. DAPI was used to visualize the nuclei. (E) Quantification by HCA of IF staining in D. Data represent mean ± SD from n = 3 (A, SA-β-Gal) and n = 4 (A [BrdU], E). (B) RNA sequencing (RNA-seq) was performed with IMR90 cells transduced with either an empty vector or ARID1B cDNA. Data were subjected to GSEA and IPA. n = 3 biological replicates were used. (C) IPA upstream regulator analysis of the RNA-seq reveals activation (z-score) of SMARCA4, TP53, and ROS pathways. (F) IMR90 cells were cotransduced with an empty vector or ARID1B cDNA and shRNAs targeting p21CIP1, p16INK4a, or p53. Cell proliferation was assessed by crystal violet staining. (***) P < 0.001; (**) P < 0.01 by two-tailed Student t-test. Bars, 30 μm.