Fig. 2: SIRT1/FOXO1 axis mediates the induction of adipose KLF10 during exercise.

a Schematic representation of Klf10 proximal promoter constructs used for luciferase assays. Predicted consensus of FOXO1-binding site is shown in the wild type (WT) luciferase construct. The red letters indicate mutations of the FOXO1-binding site in the FOXO1-Mut construct. b, c Luciferase activities were measured in HEK293T cells. Data were normalized to the vector group (n = 5 independent biological replicates). d–h Chow diet-fed 8-week-old male mice were treated as in Fig. 1a before being sacrificed for analysis. d, e FOXO1 enrichment on promoter of the indicated gene in iWAT and eWAT, respectively (n = 6 male mice/group). f FOXO1 acetylation was examined in iWAT and eWAT. Representative western blotting was shown. g, h SIRT1 activity in iWAT and eWAT, respectively (n = 6 mice/group). i Differentiated 3T3-L1 adipocytes were treated with or without SIRT1 antagonist Ex527 for 24 h (50 μM). Then Klf10 mRNA in the cells were determined (n = 6 independent biological replicates). j–l Differentiated 3T3-L1 adipocytes were treated with or without SIRT1 agonist Srt1720 for 24 h (2 μM). Then Klf10 mRNA levels, KLF10 protein levels and FOXO1 acetylation levels in the cells were determined, respectively (n = 6 independent biological replicates). m Differentiated 3T3-L1 adipocytes were treated with or without Ex527 or Srt1720 for 24 h. Then FOXO1 enrichment on promoter of the indicated gene in the cells were determined (n = 6 independent biological replicates). n Differentiated 3T3-L1 adipocytes were transfected with the siRNA against FOXO1, with siNC as the control. After 36 h, the Foxo1 mRNA levels in the cells were determined (n = 6 independent biological replicates). o Differentiated 3T3-L1 adipocytes with or without the knockdown of FOXO1 were cultured in the presence or absence of Srt1720 for 24 h. Then Klf10 mRNA in the cells were determined (n = 6 independent biological replicates). For statistical analysis, one-way ANOVA plus Tukey’s post hoc tests were performed in b, n; two-way ANOVA plus Tukey’s post hoc tests were performed in c–e, m, o; unpaired two-tailed Student’s t tests were performed in g–j. All data show the means ± SD. Source data are provided as a Source Data file.