Anti ucp1

OLZ for in vivo treatment was purchased from Hansoh Pharma (Jiangsu, China), while OLZ for in vitro experiments was purchased from Adamas-beta (Shanghai, China; H20010799). Poly I:C injections were obtained from Guangdong Bangmin Pharmaceutical Factory Co. Ltd. (Guangdong, China; H20003518). IL-6 (Wuhan, China; KA0278) and CXCL1(Wuhan, China; KA0554) enzyme linked immunosorbent assay (ELISA) kits were obtained from AmyJet Scientific. The antibodies used for immunoblotting anti-PRDM16 (1:1000, abs104818) were purchased from Absin Bioscience Inc. (Shanghai, China). The anti-phospho-AMPK (1:1000, #2531) and anti-AMPK (1:1000, #2603) were obtained from Cell Signaling Technology (Shanghai, China). Santa Cruz Biotechnology (Shanghai, China) provided the anti-PGC-1α (1:1000; sc13,067), anti-PPAR-γ (1:1000; sc-6285), anti-UCP1 (1:1000; sc-6529), anti-Actin (1:2000, sc47778), and Goat Anti-Rabbit (1:2000; sc 2005) IgG H&L (HRP, 1:5000; sc2357). The Mito-Probe 5, 50, 6, 60-tetrachloro-1, 10, 3, 30-tetraethyl-benzimidazolcarbocyanine iodide (JC-10; BB-41052) assays’ kit and Mito-Tracker Green (C1048) were obtained from BestBio Science (Shanghai, China) and Beyotime Biotechnology (Shanghai, China), respectively. All the other chemicals were acquired from Dingguo Changsheng (Beijing, China).

Immunocytochemistry was performed on formalin-fixed cells. These cells were incubated with anti-UCP1 (dilution 1:1000, Santa Cruz Biotechnology) primary antibody at 4 °C overnight, followed by incubation with appropriate FITC goat anti-mouse secondary antibody at room temperature for 4 h. For staining of mitochondria, MitoTracker^®Red (1 mM, Cell Signaling Technology, Inc.) was directly added to PBB-T (PBS + 1%, BSA, and 0.1% Tween 20) at a concentration of 200 nM. Cells were then incubated at 37 °C for 2 h. After incubation, tissues were washed with PBS and subjected to immunostaining. Morphological findings were observed using a light microscope at 40× magnification.