Fig. 4
IL-6 expression is reduced in adipocytes overexpressing Grem2. A 3T3-L1-mock or 3T3-L1-Grem2 cells were differentiated into adipocytes, and each CM was obtained from day 7 to day 9 when adipogenesis was observed. MTV/TM-011 or MDA-MB-231 cells were seeded in 96 well plates and incubated with each CM for 72 h, followed by MTS assay. B 3T3-L1 cells were differentiated using the standard induction cocktail with vehicle or GREM2 recombinant protein (50 ng/ml) for 9 days. RNA was isolated from each cell line and the expression of the indicated genes was analyzed by qPCR. C 3T3-L1-mock or 3T3-L1-Grem2 cells were differentiated using the standard induction cocktail for 9 days. RNA was isolated from each cell line and the expression of the indicated genes was analyzed by qPCR. D 3T3-L1-mock or 3T3-L1-Grem2 cells were differentiated into adipocytes, and IL-6 content was examined in each adipocyte CM obtained on the 9th day of differentiation. Two-sided t-test. *, p < 0.05; **, p < 0.01; ***, p < 0.001; ns, not significant. E Adipocytes-mock or adipocytes-Grem2 cells were treated with or without MSAB (10 μM) for 24 h. RNA was isolated from each cell line and the expression of the indicated genes was analyzed by qPCR. Two-way ANOVA. ***, p < 0.001; ns, not significant