Journal: Gastro Hep Advances
Article Title: Helicobacter pylori Exploit Short-Chain Fatty Acids-Induced CAPZA1 Overexpression to Emerge CD44v9-Positive Stemness
doi: 10.1016/j.gastha.2025.100860
Figure Lengend Snippet: SCFAs induce CAPZA1 expression in AGS cells and promote CagA accumulation and CD44v expression under H pylori infection. (A) Western blotting analysis of CAPZA1 expression in AGS cells treated with propionate (Pro) or butyrate (But), with signal quantification using ImageJ software. (B) HDAC activity in AGS cells treated with Pro or But. Statistical analysis was performed using one-way ANOVA followed by Tukey’s test. ∗ P < .05, ∗∗ P < .01. (C) ChIP assay showing histone acetylation in the CAPZA1 promoter region of AGS cells treated with Pro or But. Chromatin was immunoprecipitated with anti-histone H3 (acetyl K9) or control IgG, and enrichment of CAPZA1 promoter regions was assessed using real-time PCR. (D) Immunofluorescence analysis of autolysosome formation in But-treated AGS cells infected with H pylori ATCC 700392. Lysosomes and nuclei were stained with LysoTracker and 4ʹ,6-diamidino-2-phenylindole, respectively. Scale bar = 50 μm. (E) Western blotting analysis of CagA accumulation in But-treated AGS cells infected with H pylori ATCC 700392. (F) Western blotting analysis of CD44v expression in Pro- or But-treated AGS cells infected with H pylori G27 or G27 Δ cag PAI strains.
Article Snippet: AGS cells were infected with H pylori strains ATCC 700392, G27 ( H pylori G27), or a cag pathogenicity island ( cag PAI)-deleted isogenic mutant ( H pylori G27 Δ cag PAI) at a multiplicity of infection of 50 for 5 h. Following infection, the cells were washed with PBS and subsequently incubated in RPMI 1640 medium containing 400 μg/mL kanamycin for 24 h to kill the bacteria and prevent further injection of CagA.
Techniques: Expressing, Infection, Western Blot, Software, Activity Assay, Immunoprecipitation, Control, Real-time Polymerase Chain Reaction, Immunofluorescence, Staining
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