Fig. 2. Rapamycin specifically reduces TNF-α mRNA level. (A)RBL 2H3 cells were pretreated with either rapamycin (10 nM) or actinomycin D (10 μg/ml) for 30 min and stimulated with antigen for 15 min. (B) RBL 2H3 cells were pretreated with either wortmannin (500 mM) or LY294002 (25 μM) for 30 min and stimulated with antigen for 15 min. (C) RBL 2H3 cells were pretreated with either calphostin C (0.5 μM) or rottlerin (5 μM) for 30 min and stimulated with antigen for 15 min. (D) RBL 2H3 cells were pretreated with either EGTA (2 mM) or BAPTA-AM (25 μM) for 30 min and stimulated with antigen for 15 min. TNF-α mRNA level was analyzed by RT-PCR. GAPDH was used to normalize the amount of loaded RNA.
