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. 2016 Jan 22;7:10477. doi: 10.1038/ncomms10477

Figure 4. p38γ and p38δ interact with mTOR through DEPTOR.

Figure 4

(a) Endogenous mTOR, Raptor, Rictor, Sin-1 and DEPTOR co-immunoprecipitate with endogenous p38γ. We immunoprecipitated p38γ from WT and p38γ/δ−/− MEF lysates using specific antibodies; immunoprecipitates (IP), supernatants and total lysates were analysed by SDS–PAGE using the antibodies indicated. (b,c) p38γ interacts with mTOR through DEPTOR. HEK-293 cells were transfected with HA-p38γ, Flag-DEPTOR or Myc-mTOR or a combination of these and immunoprecipitated with the indicated antibodies targeting the c-myc epitope (b) or Flag (c). Immunoblots were probed with the indicated antibodies. (d) Co-immunoprecipitation of p38γ and p38δ with DEPTOR in HEK-293 cells. HA-p38γ or HA-p38δ expression vectors were co-expressed with Flag-DEPTOR in HEK-293T cells. Anti-Flag immunoprecipitates were analysed by SDS–PAGE. (e) HA- p38δ co-immunoprecipitates with endogenous p38γ. p38γ immunoprecipitates from HEK-293T cells transfected with HA-p38δ were analysed by SDS–PAGE. IP, immunoprecipitation; TL, total lysate.