Fig. 7

T0070907 reversed the inhibitory effect of GEN on adipocyte development to CAA and resulting inflammation in a co-culture system. (A) BODIPY 493/503 staining results and quantitative analysis of mature adipocytes cultured alone or in co-culture with E0771 cells (presence/absence of GEN and T0070907) for 4 days (n = 3). (B) Representative bright field and Pref-1 immunohistochemical staining photographs of 3T3-L1 cells. (C) Relative mRNA expression of PPAR-γ, C/EBPα, APD and FABP4 (n = 4). (D) Relative mRNA expression of Pref-1 and HSL (n = 4). (E) Immunofluorescence staining for PPAR-γ (green) and NF-κB (red) in bottom chamber 3T3-L1 cells. Nuclei were stained with DAPI (blue). (F) The protein expressions of nuclear PPAR-γ, NF-κB and cytosolic PPAR-γ, NF-κB. Histone-H3 was used as a loading control for the nuclear proteins, whereas β-actin and β-tubulin was used for the cytosolic proteins (n = 3). (G) The protein expressions of TNF-α, IL1β, IL6 and CCL-2 were measured by western blotting analysis, and β-tubulin protein level was used as a loading norma (n = 3). All values were expressed as mean ± S.D. * P < 0.05, ** P < 0.01 vs. Coculture group, # P < 0.05, ## P < 0.01 vs. Gen + Coculture group