Cd206 antibody

Samples came from 2-4-week-old C57Bl/6J mice (range of weights = 10-15 g), 5-6-week-old C57Bl/6J mice (15-20 g), and 5-6-week-old C57Bl/6J-ob/ob mice (30-35 g) and were sourced from SiBeiFu Laboratory Animal Technology Company, Beijing, China. Samples from dissections were placed into Dulbecco's modified Eagle medium (Hyclone), with additions of fetal bovine serum (Gibco), penicillin-streptomycin antibiotics, collagenase type I (Sigma), insulin (Sigma), rosiglitazone (Sigma-Aldrich), dexamethasone (Sigma-Aldrich), and isobutyl-methylxanthine (IBMX; Sigma-Aldrich). We used 24-well culture plates (Corning), 96-well culture plates (Corning), a high-fat diet (HFD), Oil-Red-O solution (abcam), paraformaldehyde (Sigma-Aldrich), isopropanol (Sigma-Aldrich), hematoxylin (Sigma), glycogelatin (Solarbio), anti-PPAR-γ (CST), anti-FABP4 (CST), anti-LPL (abcam), anti-UCP1 (CST), F4/80 antibody (BioLegend), CD206 antibody (BioLegend), SDS-PAGE assay products (beyotime), TNF-α, IL-6, IL-1β, IL-10 ELISA kits (MultiSciences), Cholesterol Assay Kit-HDL, LDL/VLDL (abcam), anti-LPA (abcam), anti-ITGAM (CST), and anti-NF-κB (CST).

Tetraethyl orthosilicate (TEOS), Cetyltrimethylammonium bromide (CTAB) were obtained from Sinopharm Chemical Reagent Co. Urea, ferric acetylacetonate (Fe(acac)₃), fluorescein isothiocyanate (FITC), Cerium (III) acetate hydrate, anhydrous dimethylsulfoxide (DMSO), 2′,7′-Dichlorofluorescin diacetate (DCF-DA) and 2,2-diphenyl-1-picrylhydrazyl (DPPH) were purchased from Sigma-Aldrich (USA). mPEG-NH2 (5kD) was purchased from Aladdin (China). Sodium salicylate (NaSal), triethanolamine (TEA), Cell Counting Kit-8 (CCK-8) were purchased from MedChemExpress (MCE, USA). Penicillin, streptomycin, fetal bovine serum (FBS), and Dulbecco’s Modified Eagle’s Medium (DMEM) were purchased from Hyclone (USA). 4′, 6-Diamidino-2-phenylindole (DAPI), Lyso-Tracker Red, total superoxide dismutase assay kit with WST-8, and catalase activity assay kit were purchased from Solarbio (China). CD68 antibody, Alexa Fluor 549 anti-Nos2 (iNOS) antibody, and CD206 antibody were purchased from Biolegend (USA). Tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β) ELISA kit were purchased from FineTest (China).

The cells were adjusted to the number of 1 × 10⁶ per sample. Use CD16/32 antibody (Biolegend, San Diego, CA, USA) to block nonspecific F_C-mediated interaction. Next, cells were incubated with CD86 antibody (Biolegend, San Diego, CA, USA) on ice for 30 min in the dark. Then cells were fixed, permeabilized, and incubated with CD206 antibody (Biolegend, San Diego, CA, USA) at room temperature for 30 min. Finally, use 500 µl PBS buffer containing 3% FBS to resuspend the cells before it being analyzed on flow cytometry (BD Biosciences, San Diego, CA, USA) [18 ].