Fig. 8
From: TBK-binding protein 1 regulates IL-15-induced autophagy and NKT cell survival
Tbkbp1 recruits PP6 to Ulk1 to oppose mTORC1-mediated Ulk1 phosphorylation. a Co-IP analysis of mTOR-Ulk1 interactions (upper panels) and direct IB analysis of the indicated proteins (lower panels) in NKT hybridoma cells transduced with a control or Tbkbp1-specific shRNA, either not treated (NT) or incubated with IL-15 or the mTORC1 inhibitor rapamycin (Rap). IgG was used as a negative control for IP. b Co-IP analysis of Tbkbp1-ANKRD28 interactions and direct IB assays using lysates of HEK293 cells transfected with the indicated expression vectors. c Co-IP analysis of ULK1 association with Tbkbp1 and ANKRD28 using lysates of HEK293 cells transfected with the indicated expression vectors. d Co-IP analysis of Ulk1 interaction with FIP200, Tbkbp1, and ANKRD28 using lysates of IL-15-stimulated NKT hybridoma cells transduced with a control or Tbkbp1-specific shRNA. IgG was used as a negative control for IP. e, f IB analysis of the indicated phosphorylated (P-) and total proteins in whole-cell lysates of IL-15-stimulated NKT hybridoma cells transduced with a nonsilencing control shRNA, two different ANKRD28-specific shRNAs (e) or two different PP6c-specific shRNAs (f). Data are representative of three independent experiments
