Anti α smooth muscle actin sma

Anti-tGFP (TA150039) was purchased from OriGene (Rockville, MD). Anti-AMPKα (#5831), anti-pAMPKα-Thr172 (#2535), Anti-AMPKα1 (#2795), Anti-AMPKα2 (#2757), anti-c-myc (#5605), anti-pc-myc-Ser62 (#13748), anti-UBC9 (#4786), anti-IL-6 (#13797), anti-NaK ATPase (#7074), anti-α-smooth muscle actin (SMA; #56856), anti-SUMO2/3 (#4971), and anti-MCP-1 (#12838) were purchased from Cell Signaling Technology Inc. (Danvers, MA). Anti-β-arrestin 2 (C16D9) rabbit mAb (Cell Signaling Technology) for the western blotting analysis, and sc-365445 (Santa Cruz Biotechnology) for the IP assay. Anti-β-arrestin 1 (D8O3J) rabbit mAb (Cell Signaling Technology). Anti-α-actin (sc-47778) and anti-SUMO1 (sc-5380) were purchased from Santa Cruz Biotechnology, Inc. (Dallas, TX). Anti-GPR120 (sc-390752) and anti-SUMO2/3 immunoprecipitation (IP) beads (#BK-162) were purchased from Cellskeleton, Inc and antibody (sc-50331) were purchased from Santa Cruz Biotechnology, Inc. GW9508 (G9797) and AICAR (A9978) were purchased from Sigma-Aldrich (St. Louis, MO). DHA (CAS6217-54-5) was purchased from Cayman Chemical Company (Ann Arbor, MI). Anti-CD68 (14-0681-80) was purchased from Affymetrix-ebioscience.

RIPA Lysis Buffer supplemented with complete EDTA-free protease inhibitor cocktail tablets (Roche Applied Science, Basel, Switzerland) and phosphatase inhibitor cocktail tablets (Sigma-Aldrich) were used to extract proteins from cells and mice kidney tissues. SDS-PAGE gels were used to load protein samples, which were then transferred to PVDF membranes. After blocking for 2 h at room temperature in 5% evaporated milk in TBS+ 0.1% Tween 20, the membranes were incubated overnight at 4°C with the appropriate primary antibodies in 5% evaporated milk in TBS+ 0.1% Tween 20. The principal antibodies utilized were as follows: anti-Thymosin β4 (ab167650, Abcam, Cambridge, UK), anti-α-smooth muscle actin (SMA) (#56856, Cell Signaling Technology, Danvers, MA, USA), IL-1β (#12703, Cell Signaling), PINK1 (#6946, Cell Signaling), anti-Tom40 (H-300, Santa Cruz Biotechnology, Santa Cruz, CA), and β-actin as a loading control (#4970, Cell Signaling). A chemiluminescent substrate was used to create the signals, which were then viewed using X-ray films.