Fig. 5: ERRα^3SA phospho-deficient mice display transcriptional reprogramming and compromised metabolic homeostasis.

a Schematic of the editing strategy for ERRα^3SA mice. Cas9 cleavage site is indicated by the red arrowhead, the single-guide RNA (sgRNA) target sequence and the protospacer adjacent motif (PAM) are marked. Homology repair template contains desired mutations (red) with the resultant residue substitutions denoted. b Total and S19-phosphorylated ERRα proteins. Each lane represents tissue from one mouse, n = 3 per group. c Immunoblot (left, each lane represents tissue from one mouse, n = 3 per group) and quantification (right) of ERRα protein levels from WT and ERRα^3SA littermates upon a fasting-refeeding transition. d Overlap and pathway analysis of DEGs identified in ND-fed ERRα^3SA mice with insulin-regulated DEGs (GSE117741⁶²) (p < 0.05, |FC| ≥ 1.20). See also Supplementary Fig. 5i. e Body weight of WT and ERRα^3SA littermates on a ND (WT, n = 41; 3SA, n = 45). Box plots show centre line as median, bounds of box as 25th and 75th percentiles, whiskers as minima and maxima. f Body weights during a 14-week HFD feeding (WT, n = 12; 3SA, n = 13). g Percentage of fat and lean mass (n = 8 per group). h Representative images of liver hematoxylin and eosin (H&E) and oil red O (ORO) staining, and epididymal WAT (eWAT) H&E staining. Scale bars, 50 μm. See Supplementary Fig. 5k, m for quantification. i Whole-body energy expenditure and j regression plot of averaged energy expenditure vs. total body mass, and k respiratory exchange ratio (WT, n = 8; 3SA, n = 7). l Respiratory exchange ratio of WT and ERRα^3SA littermates maintained on a 14-week HFD (n = 7 per group). Data are represented as means ± SEM (c, f, g, i, k, l). A generalized linear model is fitted to the data (j) and estimations are displayed by the line, with the shaded region representing the standard error. *p < 0.05, unpaired two-tailed Student’s t test (c, e–g), ANCOVA (i, j), and ANOVA (k, l). Source data are provided as a Source Data file.