Human il 8

AGS cells, from a human gastric adenocarcinoma cell line (ATCC CRL-1739), were grown in Dulbecco’s modified Eagle’s medium (DMEM) (Thermo-Fisher) supplemented with 10% heat-inactivated FBS (Gemini-Benchmark). For co-culture with H. pylori, AGS cells were seeded at 1 × 10⁵ cells/well in 24-well plates 16 h prior to infection. The day of infection, medium was removed from AGS cells and mid-log-phase (optical density at 600 nm (OD) 0.3–0.6) H. pylori resuspended in DMEM–10% FBS–20% Brucella broth was added at multiplicity of infection of 10:1. Supernatants from triplicate wells of each condition were collected at 24 hrs and assayed for the IL-8 concentration using a human IL-8 enzyme-linked immunosorbent assay (ELISA) kit according to the instructions of the manufacturer (BioLegend, San Diego, CA). IL-8 values were reported as normalized values defined as a proportion increased or decreased compared to values obtained for J99, which was included in each experimental replicate. P-values were calculated from at least two biological replicates where each isolate was assayed with triplicate wells using a one-way ANOVA with Dunnett’s corrections.

Antibodies: anti‐human LIF polyclonal antibody was from Sigma (CAT# L9277). Normal goat IgG isotype control was from R&D Systems (CAT# AB‐108‐C). Anti‐LIF monoclonal antibody D25 (Kim et al, 1992 (link)) and mouse IgG2A isotype control were from Genentech.
Small‐molecule inhibitors: Baricitinib (Apexbio Technology, CAT# A414150), cobimetinib (MedChemExpress, CAT# HY‐13064), BEZ235 (Selleckchem, CAT# S1409), Z‐VAD‐FMK (R&D Systems, CAT# FMK001), Z‐DEVD‐FMK (R&D Systems, CAT# FMK004), Q‐VD(OMe)‐OPh (Apexbio Technology, CAT# A8165), 5‐AIQ hydrochloride (Sigma, CAT# A7479), CA‐074 me (Calbiochem, CAT # 205531), CA‐074 (Tocris, CAT # 4863), and CAA0225 (Calbiochem, CAT# 219502).
Recombinant Proteins: human LIF (Sigma, CAT# SRP9001), human LIF (Biolegend, CAT# 593902), human PDGF‐AA (Peprotech, CAT# 100‐13A), human Peroxiredoxin 1 (Abcam, CAT# ab74172), human IL‐8 (Biolegend, CAT# 574202), and human VEGF 165 (R&D Systems, CAT# 293‐VE).

Media from cultured cells were collected and used for measuring IL-8 and lactate levels with Human IL-8 (Biolegend, San Diego, CA) ELISA kit^{42 (link)} and Lactate Colorimetric Assay Kit II (BioVision)^{17 (link)}.