Fig. 6: The YAP/TAZ–TEADs axis cooperates with SMAD2 to regulate fibrotic responses.
From: The Hippo pathway links adipocyte plasticity to adipose tissue fibrosis
a Representative scWAT sections with Masson’s trichrome staining of indicated mouse strains at 5 weeks old. Independent experiments were performed three times with similar results. b mRNA expression of fibrosis and adipocyte markers (n = 6 mice). Data are shown as fold change in L1L2-AKO, L1L2YT-AKO, L1L2Y-AKO, and L1L2T-AKO compared to corresponding WT littermates. The column of L1L2-AKO had also been shown in Fig. 1m. c–n Differentiated adipocytes from Cas9Tg/+ scWAT was transduced with the indicated gRNAs on day 2 post differentiation, and then analyzed 5 days later. c mRNA expression of adipocyte markers in differentiated adipocytes transduced with Vec (n = 4 biologically independent cell cultures), Lats1/2 gRNA (L1L2-gRNA) (n = 3 biologically independent cell cultures) and LATS1/2-YAP/TAZ gRNA (L1L2YT-gRNA) (n = 4 biologically independent cell cultures). d Immunoblot analysis of protein expression in cells in c. e Schematic diagram of the Lats1/2-Tead1/2/3/4 Tandem gRNA construct. f mRNA expression of adipocyte marker in differentiated adipocytes transduced with Vec, L1L2-gRNA, or Lats1/2-Tead1/2/3/4 gRNA (L1L2-Tds-gRNA) (n = 3 biologically independent cell cultures). g YAP and TAZ protein level in the nuclear/cytoplasmic fractionation of differentiated adipocytes transduced with Vec, L1L2-gRNA and L1L2-Tds-gRNA. h Immunoblot analysis of protein expression of SMAD2, YAP and TAZ in differentiated adipocytes transduced with Vec and L1L2-gRNA. i Quantification of relative density of SMAD2 normalized to HSP90 (n = 3 biologically independent cell cultures). j mRNA expression of Smad2 of cells in i (n = 3 biologically independent cell cultures). k Immunoblot analysis of SMAD2 protein expression in differentiated adipocytes (from SVF of Cas9Tg/+ scWAT) transduced with the indicated gRNAs. l Immunoblot analysis of SMAD2 protein expression in a cycloheximide chase experiment. m, Regression analysis of SMAD2 protein stability in l. n Differentiated adipocytes were transduced with MYC-Ub, L1L2-gRNA-SMAD2-HA and YT-gRNA. Immunoprecipitation (IP) assay showing an increased ubiquitination of SMAD2 in Yap/Taz-deficient cells. Data are means ± SEM. One-way ANOVA with Bonferroni’s multiple-comparisons test in c, f; two-tailed unpaired Student’s t test in b, i, j; *P < 0.05, **P < 0.01, ***P < 0.001; NS, not significant. Exact P values are provided in a Source data file.
