Fig. 4. The effects of rapamycin on TNF-α expression are not mediated by S6K1. (A) RBL 2H3 cells were pretreated with either rapamycin (10 nM) for 30 min or not 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 EGTA (2 μmM) or BAPTA-AM (25 μM) for 30 min and stimulated with antigen for 15 min. (D) 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. Cell lysates were subjected to 12% SDS-PAGE and analyzed with immunoblotting using indicated antibody.
