Anti nf κb p65

For cells, glass chambers were treated with poly‐L‐lysine for 1 h, and then gastric cancer cells were grown into the chamber (FALCON, #354118), and cells were fixed with 4% PFA for 1 h at room temperature. For tissues, the tumor samples were embedded in paraffin and the samples were cut into 4 μm sections. The sections were dewaxed in xylene and then gradually dehydrated with gradient alcohol and excess wax was removed. Sections were subjected to antigen repair in Citrate solution (10 mM, pH 6.0) at 100°C for 20 min. Next, cells or slides were treated with 0.1% Triton X‐100 for permeabilization on ice for 5 min, followed by dressing with 5% BSA for 1 h at room temperature to reduce non‐specific staining. Cells or slides were incubated overnight at 4°C with primary antibodies (anti‐NF‐κB p65, Beyotime, #AN365, 1:200 dilution; anti‐Ki67, abcam, #ab15580l, 1:500 dilution; anti‐PCNA, Dako, #M0879, 1:500 dilution; anti‐CD206, Proteintech, #18704‐1‐AP, 1:200 dilution) followed by secondary antibodies (Alexa Fluor™ 555 anti‐rabbit IgG, ThermoFisher Scientific, #A‐31572, 1:500 dilution; Alexa Fluor™ 488 anti‐mouse IgG, ThermoFisher Scientific, #A‐11029) for 1 h at 37°C. Cell nuclei were stained with DAPI (Beyotime, #C1002).

Indole-3-acetic acid and LPS (lipopolysaccharides from Escherichia coli O55:B5) were obtained from Sigma-Aldrich (St. Louis, MO, USA). Tin protoporphyrin IX (SnPP) was obtained from Cayman Chemical (Ann Arbor, MI, USA). CH-223191 was purchased from MedChem Express (Monmouth Junction, NJ, USA). Dulbecco’s modified Eagle’s medium (DMEM) was from Biological Industries (Kibbutz Beit Haemek, Israel). Fetal bovine serum (FBS) was purchased from Hyclone (South Logan, UT, USA). Cell counting kit (CCK-8), TRIzol RNA extraction reagent, nitric oxide assay kit, 2′,7′-dichlorofluorescin diacetate (DCFH-DA), and Hoechst 33,342 were obtained from Applygen Technologies Inc. (Beijing, China). Hieff^® qPCR SYBR^® Green Master Mix and Hifair^® II 1st Strand cDNA Synthesis SuperMix for qPCR (gDNA digester plus) were procured from Yeasen biotech Co. (Shanghai, China). Antibody against β-actin was from Santa Cruz Bio. (Santa Cruz, CA, USA). Anti-HO-1 was purchased from ABclonal Inc. (College Park, MD, USA). Anti-NF-κB p65 was from Beyotime (Shanghai, China). ECL Western blotting detection kit, horseradish peroxidase (HRP)-conjugated anti-rabbit immunoglobulin G (IgG), and anti-mouse IgG were purchased from Huaxingbio. (Beijing, China). ELISA kits for IL-1β, IL-6, TNF-α, and MCP-1 were bought from ExCell Biotech (Taicang, Jiangsu, China).

Sprague-Dawley (SD) rats were purchased from Vital River Laboratories Animals Technology Co., Ltd. (Beijing, China). PIO was donated by Huadong Medicine Co., Ltd. (Hangzhou, China). GW9662, diphenylene iodonium (DPI) and pyrrolidine dithiocarbamate (PDTC) were purchased from Sigma (St. Louis, MO, USA). Dulbecco’s modified Eagle medium (DMEM) and fetal bovine serum (FBS) were purchased from Hyclone Laboratories, Inc. (Logan, UT, USA). TRIzol^® reagent and cell lysates were purchased from Invitrogen Life Technologies (Shanghai, China). Anti-GAPDH was purchased from ProMab Biotechnologies Inc. (Richmond, CA, USA) Anti-RAGE was obtained from Abcam (Hong Kong, SAR, P.R. China). Anti-NF-κB p65, anti-I-κBα, fluorescein isothiocyanate (FITC)-labeled goat anti-rabbit IgG and the reactive oxygen species (ROS) assay kit were purchased from the Beyotime Institute of Biotechnology (Jiangsu, China).