Anti gapdh

Liver-pancreas tissues were isolated from normal and overfed ins-EGFP zebrafish. A rabbit polyclonal antibody to GFP (GeneTex, Irvine, CA, USA) or anti-GAPDH (AnaSpec, San Jose, CA, USA) was used and detected via chemiluminescence. Additional methods details are given in Supplementary Materials and Methods.

Whole protein lysates from mouse intestinal tissues were prepared using RIPA lysis buffer (Thermo Fisher Scientific, Rockford, IL, USA). Samples were loaded into 10% sodium dodecyl sulfate–polyacrylamide gel electrophoresis (SDS-PAGE) gels (BIO-RAD, Hercules, CA, USA), transferred to polyvinylidene difluoride membranes (Amersham Biosciences, Buckinghamshire, UK) and incubated with primary anti-K8 antibody (Troma-1, MilliporeSigma, USA,1:500), antiPGC-1α (Abcam, Cambridge, MA, USA, 1:500), anti-phospho-MLKL (p-MLKL) (CST, 1:500), anti-AKT (CST, 1:500), anti-cleaved caspase 3 (CST, 1:500), anti-GAPDH (CST, 1:1,000) and anti-cleaved-K18 (AnaSpec, Inc., Fremont, CA, USA, 1:1,000) overnight at 4°C. Membranes were incubated with secondary monoclonal goat anti-rat antibody (Immunoreagents, Inc., Raleigh, NC, USA, 1:1,000) or goat anti-rabbit antibody (CST, 1:1,000) at room temperature for 1 h. Protein bands were visualized using an ECL detection kit (Thermo Fisher Scientific, Rockford, IL, USA) according to the manufacture's protocol.