Fig. 4

GEN suppressed extensive phenotypic changes in adipocytes co-cultured with E0771 in vitro. (A) Schematic diagram of 3T3-L1 differentiation and the coculture system. (B) Changes in viability of murine preadipocytes 3T3-L1 after treatment with gradient concentrations of GEN for 4 days (n = 6). (C) BODIPY 493/503 staining results and quantitative analysis of mature adipocytes cultured alone or in co-culture with E0771 cells (presence/absence of GEN) for 4 days (n = 3). (D) Oil-red O staining results of mature adipocytes cultured alone or in co-culture with E0771 cells (presence/absence of GEN) for 4 days. (E) Representative bright field and Pref-1 immunohistochemical staining photographs of 3T3-L1 cells. (F) Relative mRNA expression of PPAR-γ, C/EBPα, APD and FABP4 (n = 4). (G) Relative mRNA expression of Pref-1 and HSL (n = 4). (H) Relative mRNA expression of TNF-α, IL1β, IL6 and CCL-2 (n = 4). All values were expressed as mean ± S.D. ** P < 0.01 vs. Non-coculture group, # P < 0.05, ## P < 0.01 vs. Coculture group