Validation of roles of regulators and targets identified through upstream regulator analysis in cellular senescence induced by ATF6α ectopic expression.

MCF-7 cells were co-transfected with ATF6α cDNA, and control siRNA (SC) or siRNA for the indicated genes. After 5 days, SA-β-gal staining was performed. (A, C, E, G) Light microscopic images represent three independent experiments. (B, D, F, H) Percentages of SA-β-gal (+) cells were plotted. (I, K, M) Light microscopic images represent three independent experiments. (J, L, N) Percentages of SA-β-gal (+) cells were plotted. Data represent the mean of triplicate determinations ± S.D. (O, P) MCF-7 cells were transfected with ATF6α cDNA or an empty vector and treated with 100 nM Ravoxertinib, an ERK inhibitor, or 500 nM Trametinib, a MEK1/2 inhibitor. After 5 days, SA-β-gal staining was performed. Data represent the mean of triplicate determinations ± S.D. Ctrl, control; ATF6 O/E, ATF6α overexpression. * P<0.05; ** P<0.01; *** P<0.001; ns, non-significant.
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