Fig 1: Rosiglitazone alleviates lipolysis and inflammatory response.(A) Relative expression of the Atgl and Lipe (Hsl) mRNAs in isolated dWAT on day 5. n = 3 per group. (B) Western blot images and quantifications of lipase level from isolated dWAT cells at Grade 1. n = 3. (C) Transmission electron microscopy images of dermal adipocytes from Ctrl- and Afa-treated rat skin on day 5. Asterisks show small lipid droplets. Scale bars: 5 µm. (D and E) Representative images of CD68 immunostaining (D) and quantification (E) in skin from Ctrl- and Afa-treated rats at indicated times. Scale bars: 200 µm. (F) Western blot images and quantifications showing lipase levels in dFB-derived adipocytes stimulated by rat skin homogenates. n = 3 per group. (G) Relative expression of the IL6 and TNFa mRNAs in HaCaT cells after 4 hr of treatment with 100 nM Afa. n = 3 per group. (H) Schematic diagram of the experimental setup used to evaluate the roles of Rosi and adipocytes. HaCaT cells were treated with drugs for 24 h, then the supernatants were transferred directly to dFBs for another 24 hr incubation. For transwell assay, the supernatants of dFBs were added to the low chamber. (I) Confocal microscopic images of differentiated-dFBs after treatments of HaCaT supernatants from Ctrl, Afa and Afa +Rosi. Lipids were stained with BODIPY 493/503. Scale bars: 50 µm. (J and K) Representative images (J) and quantification (K) of migrated THP-1 cells stimulated with culture media from HaCaT supernatants (Ctrl, Afa, Afa +Rosi)-treated dFB cells. (L) Confocal microscopic images of HaCaT supernatant (Afa)-treated dFBs after treatment with Ctrl or Rosi. Scale bars: 50 µm. (M and N) Representative images (M) and quantification (N) of migrated THP-1 cells stimulated with culture media from HaCaT supernatant (Afa)-treated dFB cells. Then, 5 µM Rosi or vehicle was added to dFBs. (O and P) Representative images (O) and quantification (P) of migrated THP-1 cells stimulated with culture media from HaCaT supernatant (Afa)-treated undifferentiated or differentiated dFB cells. Data are presented as the means ± SEM. *p < 0.05, **p < 0.01, and ***p < 0.001 (Afa/Rosi group vs Ctrl group and Afa +Rosi group vs Afa group) using two-tailed unpaired Student’s t test and one-way ANOVA. Figure 4—source data 1.Original western blot data for Figure 4B. Figure 4—source data 2.Original western blot data for Figure 4F.