Goat anti mouse igg hrp

Cells were cultured in six‐well plates with SBSGL at 0, 600, 1200 and 1800 μg/mL concentrations. Radio immunoprecipitation assay buffer (Shanghai Biyuntian Biotechnology Co., lot no. P0013C) with a protease inhibitor was used to extract proteins. Equal amounts of protein from each sample were transferred to polyvinylidene fluoride membranes (Merck, lot no. IPFL00010). Membranes were incubated with primary antibodies against RACK1 (Cell Signaling Technology, lot no. 5432, 1:1000), OGT (Cell Signaling Technology, lot no. 24083, 1:1000), O‐GlcNAc (Thermo Fisher Scientific, lot no. MA1‐072, 1:1000), LC3B (Cell Signaling Technology, lot no. 3868, 1:1000), p62 (Cell Signaling Technology, lot no. 5114, 1:1000), PD‐L1 (Cell Signaling Technology, lot no. 13684, 1:1000), Myc‐tag (Cell Signaling Technology, lot no. 2276, 1:1000) and glyceraldehyde‐3‐phosphate dehydrogenase (GAPDH, Cell Signaling Technology, lot no. 5174, 1:1000) overnight at 4°C. After washing with Tris‐buffered saline with Tween (TBST, Biosharp, lot no. 21259419), secondary antibodies goat anti‐rabbit IgG‐HRP (Abbkine, lot no. A21020, 1:10,000) and goat anti‐mouse IgG‐HRP (Abbkine, lot no. A21010, 1:10,000) were applied for 1 h at room temperature. Membranes were washed thrice with TBST, and visualization was achieved using an ECL luminescence kit (Shanghai Biyuntian Biotechnology Co., lot no. P0018S).

LPS (Escherichia coli serotype O55:B5, L2880) was obtained from Sigma-Aldrich (USA). F4/80 (cat no. 29414-1-AP) and myeloperoxidase (MPO, cat no. 22225-1-AP) antibodies were from Proteintech (Wuhan, China), and hypoxia-inducible factor-1α (HIF-1α) antibody (cat no. BF8002) was from Affinity Biologicals (Jiangsu, China). XO (cat no. sc-398548) antibody was from Santa Cruz (USA). The XO activity assay kit (cat no. KTB1070), goat anti-mouse IgG H&L (DyLight 649) (cat no. A23610), goat anti-mouse IgG H&L (DyLight 488) (cat no. A23210), and goat anti-mouse IgG HRP (cat no. A21010) were purchased from Abbkine (Wuhan, China). Pimonidazole HCl and anti-pimonidazole mouse antibody was purchased from Hypoxyprobe (USA). ELISA kits for tumor necrosis factor-α (TNF-α, cat no. BMS607-3), interleukin-6 (IL-6, cat no. KMC0061), and IL-1β (cat no. KMC0061) were from Thermo Fisher Scientific (USA). The fluorescein (FITC) TUNEL Cell Apoptosis Detection Kit was from Servicebio (cat no. G1501, Wuhan, China). Dihydroethidium (DHE) for probing superoxide radicals was purchased from Beyotime (cat no. S0063, Shanghai, China).