Fig. 7: KLF10 forms a complex with FOXO1 and KDM4A to promote the transactivation of ATGL. | Nature Communications

Fig. 7: KLF10 forms a complex with FOXO1 and KDM4A to promote the transactivation of ATGL.

From: Exercise-induced anti-obesity effects in male mice generated by a FOXO1-KLF10 reinforcing loop promoting adipose lipolysis

Fig. 7

a Schematic representation of Pnpla2 proximal promoter constructs used for luciferase assays. The predicted consensus of FOXO1-binding site is shown in the WT luciferase construct. The red letters indicate mutations of the FOXO1-binding site in the FOXO1-Mut construct. b Luciferase activities were measured in HEK293T cells (n = 5 independent biological replicates). Data were normalized to the Vector group. c Differentiated C3H10T1/2 adipocytes were transfected with the indicated siRNAs. Then KLF10 enrichment on the promoter of Pnpla2 gene was determined. Data were normalized to the corresponding IgG control group. d, e Representative immunoblot of exogenous co-immunoprecipitation (Co-IP) in HEK293T cells and endogenous Co-IP in C3H10T1/2 adipocytes. WCL, whole cell lysates. f Enrichment of KLF10-interacting FOXO1 on promoter of the indicated gene by using the Re-ChIP method. g Enrichment of H3K9me3, H3K4me3, H3K27me3, H3K36me3 on promoter of the indicated gene in KLF10 knockout mice primary adipocytes. h C3H10T1/2 adipocytes were transfected with the indicated siRNAs and the mRNA level of Pnpla2 in the cells were examined. Data were normalized to the siNC group. i H3K9me3 enrichment on promoter of the indicated gene in C3H10T1/2 adipocytes. j Representative immunoblot of endogenous Co-IP in differentiated mice primary adipocytes with KLF10 knockout (AD-Cre) or overexpression (AD-KLF10). k, l Quantification of immunoblot results in j. Data were normalized to the AD-LacZ/Anti-FOXO1 group. m, n Differentiated primary adipocytes were infected with adenoviruses (AD) harboring the indicated genes. Then FOXO1 and KDM4A enrichment on promoter of the indicated gene in the cells was examined. n = 6 independent biological replicates in c–n. For statistical analysis, two-way ANOVA plus Tukey’s post hoc tests were performed in b, c, g, i, k–n; two-tailed Wilcoxon test was performed in f; one-way ANOVA plus Tukey’s post hoc tests were performed in h. All data show the means ± SD. Source data are provided as a Source Data file.

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