Fig. 2

Gene silencing of ERRα affects p53 biology leading to mitochondrial dysfunction and less proliferation. a HCT-116^p53+/+ (left), DLD-1 (middle), and HCT-15 (right). IB analysis was conducted using anti-ERRα, anti-p53, anti-actin, and anti-HSP-70. b IB analysis was conducted in HCT-116^p53+/+ cells using the anti-total OxPhos WB antibody cocktail, anti-PGC-1α, or HSP-90. c, d Mitochondrial DNA copy number analysis and the intracellular ATP levels were measured in HCT-116^53+/+ cells as described in the “Methods” section. e Immunofluorescence (IF) analysis for COX-4 and VDAC1 was conducted in HCT-116^53+/+ cells. Panels represent selected digitally enlarged portions of parent images to enhance the visibility of COX-4 and VDAC1. Co-localization of COX-4 and VDAC1 was quantified (as % overlay). Scale bar, 50 μm. f HCT-116^p53+/+ cell growth was analyzed. All cells were stably transduced with lentiviral constructs expressing an shRNA specific to ERRα (shERRα#) or an shRNA non-targeting construct (shMock). The data are shown as means ± S.D. (n = 2–4). The p value was calculated using a two-tailed Student’s t test. *p ≤ 0.05; **p ≤ 0.01; ***p ≤ 0.001; ****p ≤ 0.0001; n.s., not significant