Figure 6
From: TAK1 regulates autophagic cell death by suppressing the phosphorylation of p70 S6 kinase 1
TAK1 competes with S6K1 for binding to raptor.
(A) HA-raptor and Flag-TAK1 were co-transfected into HEK 293T cells. An HA immunoprecipitate was obtained and analyzed for the presence of raptor-TAK1 complexes using a Flag antibody. The blots in the lower three panels were obtained from the same cell lysates using antibodies against Flag, HA or β-actin. (B) HEK 293T cells were transfected with the indicated genes and 48 h after transfection, cell lysates were immunoprecipitated with an anti-Myc antibody, followed by immunoblotting with antibodies to HA, Flag or Myc. The blots in the lower four panels were obtained from the same cell lysates using the indicated antibodies. (C) Dose-dependent co-immunoprecipitations were performed in HEK 293T cells. Cells were transfected with the indicated constructs, along with increasing amounts of HA-raptor and incubated for 48 h. Cell lysates were collected and immunoblotted with the indicated antibodies. (D) TAK1 induces AMPK-independent autophagy. HEK 293T cells were transfected with the indicated siRNA and TAK1. The expression of TAK1, GFP-LC3-I, GFP-LC3-II, pAMPK-α, AMPK-α and β-actin were analyzed by immunoblot analysis. (‘+’ denotes 50 nM AMPK siRNA and ‘++’ denotes 100 nM AMPK siRNA.).
