Figure 4

TNFα downregulates Tmem182 transcript expression in 3T3-L1 adipocytes. After serum-starvation for 6 h, 3T3-L1 adipocytes were incubated with either DMSO vehicle (V), 50 μM PD98059 (PD), 20 μM SB20358 (SB), 50 μM LY294002 (LY), 100 nM wortmannin (WM), or 1 μM rapamycin (Rap) for 1 h and then either treated or untreated with 10 ng/ml of TNFα for 16 h. RNA samples were harvested and Tmem182 transcript level was analyzed by real-time PCR as described in Methods. Values were normalized to Gapdh transcript expression level and transcript expression in vehicle alone was set to a value of 1. Statistical analysis was carried out using one-way ANOVA followed by Tukey post-hoc test; * indicates p < 0.001 for indicated samples vs. V alone (first column). Treatment with inhibitors alone failed to appreciably alter expression for Tmem182 transcript (data not shown).