Fig. 5.

SLC15A4 is required for mTOR activity in mast cells. (A) The mTOR pathway in BMMCs. Whole-cell extracts from WT or Slc15a4^−/− BMMCs were analyzed by immunoblotting. Band intensities were measured by ImageJ software (NIH) and the relative phosphorylation ratio was indicated. (B–E) Pharmacological mTOR inhibition in IL-33 signaling in BMMCs. The mTOR signaling pathway during IL-33 stimulation (B) and levels of pro-inflammatory cytokines produced by WT BMMCs stimulated with recombinant IL-33 for 6 h in the presence of Torin 1 at the indicated concentrations (C–E). **P < 0.01; n.s., not significant. (F) Effect of pharmacological mTOR inhibition in FcεRI-triggered cytokine production in BMMCs. The levels of pro-inflammatory cytokines produced by IgE-sensitized WT BMMCs stimulated with TNP₄-BSA for 6 h in the presence of Torin 1 at the indicated concentrations determined by ELISA (in triplicate). Results of both immunoblotting (A and B) and ELISA (C–F) and results are representative of three independent experiments.