Gpr120

P38, Gαq, GPR120, GPR40, RARα, RARβ, and RARγ siRNA oligos were obtained from Gene Pharma (Suzhou, China) and used as non-targeting controls. Cells were transfected with appropriate siRNAs using jetPRIME reagent (Polyplus) according to the manufacturer’s protocol. Cells were harvested, washed twice with ice-cold phosphate-buffered saline (PBS), and subjected to western blot analysis, each western blot was measured two times as described previously [51 (link)]. The antibodies used were as follows. LC3B(#L7543) was purchased from Sigma-Aldrich. mTOR(#A2445), phospho-mTOR(S2448,#AP0094), Beclin-1(#A7353), and UVRAG(#A8462) were purchased fromAbclonal. phospho-p70S6-kinase(T389,#9234), phospho-p38(Thr180/Tyr182,#4511), phospho-ERK1/2(Thr202/Tyr204,#4370), p38(#8690), ERK1/2(#4695), and Gαq(#14373) were purchased from Cell Signaling Technology. RARα(C20,#sc-551), RARβ(C19,#sc-552), RARγ(C19,#sc-550), GPR40(FL300, sc-32905) and β-actin (N21,#sc-130656)were purchased from Santa Cruz Biotechnology. GPR120(#NBP1-00858) was purchased from NovusBiologicals. Ki67(#ab15580) and Flotilin-1 (ab133497) were purchased from Abcam. Goat anti-rabbit (A00098) secondary antibody was purchased from Genescript.

Cells were lysed with lysis buffer (50 mM Tris-HCl, 150 mM NaCl, 1% Nonidet P-40, 0.1% SDS, protease inhibitor cocktail, 50 mM NaF, and 0.2M Na₃VO₄). The protein concentration in the supernatants was measured using the Bradford assay (Bio-Rad, Hercules, CA, USA). Proteins extracts were separated on a 12% SDS polyacrylamide gel and transferred onto a nitrocellulose membrane. After blocking with 5% skimmed milk in phosphate-buffered saline with Tween-20 (PBST) for 1 h at room temperature, the membranes were probed with anti-UCP1 (#ab155117, Abcam, Cambridge, MA, USA), anti-ANP (sc515701, Santa Cruz Biotechnology, Santa Cruz, CA, USA), anti- PGC1α (#M00236, Bosterbio, Pleasanton, CA, USA), anti-Corin (#PA5-42916, Novus, Littleton, CO, USA), GPR120 (#NBP1-00858, Novus, Littleton, CO, USA) and anti-actin (#A2060, Sigma-Aldrich, St. Louis, MO, USA). After washing, the protein extracts were incubated with horseradish peroxidase-conjugated secondary antibodies and detected using an enhanced chemiluminescence substrate (Amersham Biosciences, Freiburg, Germany).