Figure 5.

Identification of factors in mast cell CM that induce adipocyte UCP1 expression. A: TIB64 mast cells were incubated at 37°C (control) or 30°C (cold) for 4 h in the absence (control) or presence of nedocromil, and the media were harvested (0 h). Another set of 30°C-incubated mast cells was allowed to recover at 37°C for 4 h, and the media were harvested (time course shown above the graph). Shown are the histamine and IL-4 protein levels measured in the media. B: Mast cell CM from A was warmed to 37°C and applied to differentiated 3T3-L1 adipocytes for 4 h at 37°C, and UCP1 mRNA was measured. C: Differentiated 3T3-L1 adipocytes in six-well dishes were cocultured with mast cells in inserts as indicated or were cultured alone (control). Nedocromil was added as indicated. The coculture system was incubated at 30°C or 37°C, and the adipocytes were harvested after 4 h of coculture at 37°C or 0 or 4 h after incubating the coculture at 30°C for 4 h (time course shown above the graph). UCP1 mRNA expression was measured in the adipocytes. Data are mean ± SEM (n = 3). *P < 0.05, **P < 0.01, ***P < 0.001 (ANOVA with a Tukey post hoc test). C, control; Ned, nedocromil.