Western primary antibody dilution buffer

MC38, a mouse colon cancer cell line, was purchased from the American Type Culture Collection (Rockville, MD, USA) and grown in DMEM supplemented with 100 ml/l fetal bovine serum under a humidified atmosphere of 5% CO₂ at 37°C. The other reagents were DMEM/HIGH GLUCOSE (HyClone; SH30022.01), fetal bovine serum (Lonsera; S711–001 s), Western Primary Antibody Dilution Buffer (Beyotime; P0023A), Super ECL Plus Western Blotting Substrate (Baoguang Biotechnology Co. Ltd BG0001), TrypLE™ Express (Thermo; 12 604 013), 10% formalin (Chongqing Chuandong Chemical), paraformaldehyde (Thermo; 305 362), EDTA (Beyotime ST066), nonfat dry milk (Cell Signaling 9999S), goat serum (Bioss; C-0005), TRIzol reagent (Invitrogen; 269 201), bovine serum albumin (BSA) (Beyotime; ST023-200 g), permeabilization buffer 10X (Invitrogen; 2 178 649), advanced D-MEM/F-12 (1X), liquid (Gibco; 12 634–010), IntestiCult™ organoid growth medium (mouse) (STEMCEL; 06000) and penicillin streptomycin solution (100X) (Beyotime; C0222).

The following reagents were purchased: lipofectamine RNAiMAX (cat. no. 13778100; Thermo Fisher Scientific, Inc., Waltham, MA, USA); opti-MEM (cat. no. 31985062; Gibco, Eggenstein, Germany); STING small interfering RNA (siRNA), negative control (NC)-siRNA (GenePharma Co., Ltd., Shanghai, China); STING small-molecule inhibitor, C-176 (cat. no. HY-112906; Med-ChemExpress, Princeton, NJ, USA); acid phosphatase, leukocyte (tartrate-resistant acid phosphatase; TRAP) Kit (cat. no. 387A-1KT; Sigma-Aldrich, St. Louis, MO, USA); mouse cytokine array (cat. no. ARY028; R&D Systems, Inc., Minneapolis, MN, USA).
The following reagents were purchased from Beyotime Institute of Biotechnology: Western Primary Antibody Dilution Buffer (cat. no. P0023A), Western Secondary Antibody Dilution Buffer (cat. no. P0023D), BeyoECL Star Super Sensitive ECL Chemiluminescence Kit (cat. no. P0018AS), BCA Protein Assay Kit (Cat. no. P0012) and Senescence β-Galactosidase Staining Kit (cat. no. C0602).