(A) qPCR analysis for the mRNA levels of Hnf4α and its target genes including G6pc, Pck1, Apoc3, and Cyp7a1 in the liver of WT and FKO mice fed on HFD (n = 5–10 per group, each point represents a biology replicate, representative of three repeats). Data are represented as mean ± SEM, one-way ANOVA, *P < 0.05. (B) WB analysis of HNF4α in the lysates from the liver of WT and FKO mice fed on HFD. Tubulin-α was used as loading control (n = 4 per group, representative of three repeats). (C) qPCR analysis for the mRNA levels of Hnf4α target genes including Pck1, Cyp7a1, and Apoc3 in the primary hepatocytes of WT or liver-specific Hnf4 knockout mice treated by the serum from WT or FKO mice (n = 3–6 per group, each point represents a biology replicate, representative of three repeats). Data are represented as mean ± SEM, t test, *P < 0.05, n.s, no significance. (D) Analysis of luciferase reporter activity for HNF4α in HepG2 cells treated by the serum from WT or FKO mice. RLU, relative light units (n = 6 per group, each point represents a biology replicate, representative of three repeats). Data are represented as mean ± SEM, t test, **P < 0.01. (E) qPCR analysis for the mRNA levels of Srebf1, Srebf2, Mlxipl, and Nr1h3 in the liver of WT and FKO mice fed on regular chow or HFD (n = 5 per group, each point represents a biology replicate, representative of three repeats) Data are represented as mean ± SEM, one-way ANOVA. (F) qPCR analysis for Creb1 mRNA in the sWAT of WT and FKO mice fed on regular chow or HFD (n = 5 per group, representative of three repeats). Data are represented as mean ± SEM, one-way ANOVA. (G) WB analysis of pCREB and CREB in the lysates from the sWAT of WT and FKO mice fed on HFD. β-Actin was used as loading control (n = 3 per group, representative of three repeats). (G, H) Quantification of the band intensity for pCREB/CREB ratio in (G) (n = 3 per group, each point represents a biology replicate). Data are represented as mean ± SEM, t test, *P < 0.05. (I) WB analysis of pCREB and CREB in the lysates from the BAT of WT and FKO mice fed on HFD. β-Actin was used as loading control (n = 4 per group, representative of three repeats). (I, J) Quantification of the band intensity for pCREB/CREB ratio in (I) (n = 4 per group, each point represents a biology replicate). Data are represented as mean ± SEM, t test, *P < 0.05. (K) WB analysis of pCREB and CREB in the lysates from the liver of WT and FKO mice fed on HFD. β-Actin was used as loading control (n = 4 per group, representative of three repeats). (K, L) Quantification of the band intensity for pCREB/CREB ratio in (K) (n = 4 per group, each point represents a biology replicate). Data are represented as mean ± SEM, t test, **P < 0.01.