Extended Data Fig. 7: Identification of HSL phosphorylation site by PAK4.
From: p21-activated kinase 4 counteracts PKA-dependent lipolysis by phosphorylating FABP4 and HSL
a. After transfecting 3T3-L1 adipocytes as indicated, co-IP was performed to determine PAK4′s interaction with and phosphorylation of ATGL. b. 3T3-L1 cells were transfected with PAK4 and HSL. Phosphorylation of HSL was determined using LC-MS/MS analysis after in-gel digestion. A representative spectrum is shown. c. The primary sequence of murine HSL is shown with highlighted sequence coverage (78%; letters in bold) and phosphorylation of HSL (S565, letter in red) revealed by LC-MS/MS. d. 3T3-L1 adipocytes were transfected as indicated and then treated with 1 μM isoproterenol (Iso) for 1 h. Phosphorylation of HSL, glycogen synthase kinase-3 beta (GSK3β), and AMP activated kinase (AMPK) was determined by Western blotting. Data are representative of at least three independent experiments. Source data are provided as a Source Data file.
