Fig. 4: Mitochondrial master regulator Estrogen-Related Receptor alpha mediates PMVK-induced senescence.
From: Cholesterol biosynthetic pathway induces cellular senescence through ERRα
a, b RT-qPCR of ESRRA and ERRα target genes (including UQCRFS1, NDUF5A, SDHA, SDHB) in empty vector (Ctrl), PMVK- or Kinase Dead (PMVKmut) PMVK-expressing cells. Mean +/− SEM of n = 4 independent biological replicates. RM one-way ANOVA test. c, d RT-qPCR of ESRRA and ERRα target genes in Ctrl and PMVK-expressing cells, upon siCtrl or siERRα transfection. Mean +/− SEM of n = 8 independent biological replicates. RM one-way ANOVA test. e Representative micrographs and cell number quantification of Ctrl and PMVK-expressing cells previously transfected with siERRα. Scale bar: 100 µm. Mean +/− SEM of n = 5 independent biological replicates. RM one-way ANOVA test. f Crystal violet staining of Ctrl and PMVK-expressing cells previously transfected with siERRα. g Quantification of SA-β-gal positive cells in Ctrl and PMVK-expressing cells upon siERRα transfection. Mean +/− SEM of n = 5 independent biological replicates. RM one-way ANOVA test. h RT-qPCR of p21CIP1 and IL-8 genes in Ctrl and PMVK-expressing cells previously transfected with siCtrl or siERRα. Mean +/− SEM of n = 7 independent biological replicates. RM one-way ANOVA test. i Western blot on ERRα, p21CIP1 and Tubulin in liver of ERRα WT and ERRα KO male mice fed either by chow diet (CD) or high-fat diet (HFD). Quantification of p21CIP1 levels normalized to Tubulin levels. Mean +/− SEM of n = 3–4 male mice. Ordinary one-way ANOVA test. j, k RT-qPCR of p53 target genes (namely p21Cip1, Gadd45a, Gdf15) and SASP members Cxcl1, Cxcl2 and Mmp12 genes in liver of ERRα WT and ERRα KO male mice fed either by chow diet (CD) or high-fat diet (HFD). Mean +/− SEM of n = 3–4 mice. Ordinary two-way ANOVA test. (*p < 0.05; **p < 0.01; ***p < 0.001).
