Fig. 2: TGFβ stimulation is coupled with Hippo pathway inactivation to promote AT fibrosis.

a Quantification of phosphorylation levels of SMAD2 in scWAT during growth (n = 4 mice). b Eight-week-old male L1L2-FF or Lats1^f/fLats2^f/fAdipoq^CreERT2 (L1L2-iAKO) mice were locally injected with AAV-CAG-GFP and AAV-CAG-TGFβR1 (T204D) respectively in scWAT. After 3 days, all mice were intraperitoneally (i.p.) administered 3 doses of tamoxifen every other day and then analyzed 4 weeks later. c Immunostaining of αSMA⁺ cells in the indicated mice. Independent experiments were performed twice with similar results. d mRNA expression of fibrotic and adipocyte markers in L1L2-FF or L1L2-iAKO scWAT transduced with AAV-CAG-GFP (n = 5 mice) or AAV-CAG-TGFβR1 (T204D) (n = 6 mice) followed by tamoxifen administration. e, f L1L2-FF or L1L2-iAKO mice were locally injected with AAV-CAG-TGFβ1 (2CS) in scWAT. e mRNA expression of fibrotic and adipocyte markers (n = 5 mice). f Immunostaining of αSMA⁺ cells. g AAV vectors for inducible expression of TGFβR1 (T204D) by Double-Floxed Inverted Open reading frame system. h, i, mRNA expression of fibrotic and adipocyte markers (h) (n = 4 mice) or immunostaining of αSMA⁺ cells (i) in L1L2-FF or L1L2-iAKO scWAT transduced with AAV-CAG-GFP or AAV-EF1A-DIO-TGFβR1. j Eight-week-old male L1L2-FF and L1L2-iAKO mice were subcutaneously co-injected with AAV-ADP-FLPo and AAV-EF1A-FIO-TGFβR1 (T204D) for 3 weeks, followed by i.p. injection of tamoxifen. k, l mRNA expression of fibrosis markers (k) (n = 6 mice) or immunostaining of αSMA⁺ cells (l) of mice in j. m mRNA expression of Acta2 of Cas9^Tg/+ SVF transduced with vector (n = 4 biologically independent cell cultures) or YT-gRNA (n = 3 biologically independent cell cultures) in the presence or absence of TGFβ1 for 36 h. Data are means ± SEM. Two-way analysis of variance (ANOVA) with Bonferroni’s multiple-comparisons test in d, e, h, k, m; *P < 0.05, **P < 0.01, ***P < 0.001; NS, not significant. Exact P values are provided in a Source data file.