Atg14

The WES capillary electrophoresis system (ProteinSimple, San Jose, CA) was used following standard manufacturer protocols to assess protein levels on total liver lysates. Compass for SW (v4.0.0; ProteinSimple) was used to assess the area under each peak using the Gaussian fit method. The following antibodies and dilutions were used from the respective manufacturers: PDH (1:50; #2784; Cell Signaling, Danvers, MA); PDH p-Ser300 (1:50; AP1064; Sigma-Aldrich); ACLY (1:65; #4332; Cell Signaling); Total Acetylated Lysine (1:65; #9441; Cell Signaling); AMPK (1:50; #2603; Cell Signaling); AMPK p-Thr172 (1:50; #2535; Cell Signaling); ATG14 (1:50; PD026MS; MBL International, Woburn, MA); ATG14 p-Ser29 (1:50; #92340; Cell Signaling).

Phosphospecific antibodies were as described: pT172 (AMPK-α: Cell Signaling Technology, Danvers, MA, USA, Cat# 2535, RRID AB_331250), pS79/pS212 (ACC1/ACC2: Cell Signaling Technology, Cat# 11818, RRID AB_2687505), pS792 (Raptor: Cell Signaling Technology, Cat# 2089146, RRID AB_2934061), pT1337 (GBF1: IBL International, Hamburg, Germany; Cat# 28065-IBL, RRID AB_2232232), and pS29 (ATG14: Cell Signaling Technology, Cat# 92340, RRID AB_2800182). Antibodies against total proteins were as described: AMPK-α1 and AMPK-α2 (for immunoprecipitation): [49 (link)]; AMPK-α (pan-α, for Western blotting): Cell Signaling Technology, Cat#2793, RRID AB_915794GBF1; Raptor: Cell Signaling Technology, Cat# 2280, RRID AB_561245; GBF1: BD Biosciences, San Jose, CA, USA, Cat# 28065-IBL, RRID AB_2232232; ATG14: MBL International, Woburn, MA, USA, Cat# PD026, RRID AB_1953054; tubulin: Sigma-Aldrich, St. Louis, MO, USA, Cat# T4026, RRID AB_477577; α-actin: Sigma-Aldrich, Cat# A2228, RRID AB_476697. Streptavidin conjugated to 800 nm fluorophore (for the detection of total ACC) was from Rockland Immunochemicals, Pottstown, PA, USA, Cat# S000-32.