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. 2021 Jun 22;10:e69209. doi: 10.7554/eLife.69209

Figure 7. Adipsin is a downstream target of PPARγ deacetylation.

(A) Immunoblot of Adipsin in the plasma from adult male adipocyte conditional PPARγ KO (Adipoq-Pparγ KO) and control mice on HFD for 12 weeks (L.C. = Coomassie staining of the membrane). (B) Adult male WT and 2KR mice on HFD for 12 weeks followed by 8 weeks of HFD or HFD supplemented with rosiglitazone (Rosi). Immunoblots of Adipsin, Adiponectin (APN), and aP2 from epididymal white adipose tissue (EWAT) (loading control = cyclophilin A) and Adipsin and APN from plasma (L.C. = Coomassie stain of the membrane). (C) Adipsin promoter-driven luciferase reporter assay from HEK293T cells transfected with WT or 2KR overexpression of PPARγ with or without Rosi treatment (n = 3/group). (D) ChIP assay for PPARγ binding to the Adipsin promoter. Pparg-/- mouse embryonic fibroblasts (MEFs) were reconstituted with Flag-HA-tagged WT or 2KR PPARγ2 and adipogenesis was induced. Anti-HA ChIP assay was performed on day 7 of differentiation. (E) Scheme of Adipsin promoter designs: long Adipsin promoter (APL), short Adipsin promoter (APS), Adipsin promoter core (APC), delete −197 to −183 (dP1), delete −164 to −151 (dP2), delete −165 to −50 (dP3). (F, G) Adipsin promoter-driven luciferase reporter assay in HEK293T cells with various deletions in the Adipsin promoter region. ***p<0.001 for WT vs. 2KR. Data represent mean ± SEM. Two-tailed Student’s t-tests were used for statistical analyses.

Figure 7.

Figure 7—figure supplement 1. PPARγ deacetylation represses Adipsin expression in peripheral adipose tissues.

Figure 7—figure supplement 1.

(A–C) Adult male WT and 2KR mice on HFD for 12 weeks followed by 8 weeks of HFD supplemented with rosiglitazone (Rosi). qPCR analyses of Cfd expression in epididymal white adipose tissue (EWAT) (n = 9, 9) (A), subcutaneous white adipose tissue (SWAT) (n = 9, 9) (B), brown adipose tissue (BAT) (n = 6, 6) (C). (D) qPCR analysis of Cfd expression in the bond marrow (BM) of ad libitum-fed adult male WT and 2KR mice (n = 5, 6). *p<0.05, **p<0.01, ***p<0.001 for WT vs. 2KR. Data represent mean ± SEM. Two-tailed Student’s t-tests were used for statistical analyses.