Additional file 2 of Helicobacter pylori disrupts gastric mucosal homeostasis by stimulating macrophages to secrete CCL3
Creators: name: Wei, Yan-Fei
name: Li, Xue
name: Zhao, Meng-Ran
name: Liu, Si
name: Min, Li
name: Zhu, Sheng-Tao
name: Zhang, Shu-Tian
name: Xie, Si-An
Publisher: name: figshare
Year: 2024
Subjects: subject: Biochemistry
subject: Space Science
subject: Medicine
subject: Microbiology
subject: FOS: Biological sciences
subject: Chemical Sciences not elsewhere classified
subject: Science Policy
subject: Immunology
subject: FOS: Clinical medicine
subject: Cancer
subject: Infectious Diseases
subject: FOS: Health sciences
subject: Computational Biology

description: Additional file 2: Supplementary fig. 1 H. pylori-infected macrophage medium disrupts gastric mucosal barrier. A TEER of MKN28 cells with different culture media. B-C The protein level of tight junction in MKN28 and GES-1 with different culture media. D IF detected the tight junction between MKN28 cells with different culture media. DAPI was used for nuclear staining (blue), Occludin was used for tight junction staining (green), arrows showed the disruption of tight junctions between cells. Abbreviations: NC, normal culture medium; MC, macrophage culture medium; HMC, H. pylori-infected macrophage culture medium; IF, Immunofluorescence. The data are presented as the mean±S.D. after triplicate. Two groups were compared by t-test, multiple groups were compared by one-way analysis of variance (ANOVA). Note: **P &lt; 0.01. Supplementary fig. 2 H. pylori stimulates macrophages to secrete chemokines. A Analysis chemokines in the medium of THP-1 co-cultured with H. pylori by antibody chip. B Differential analysis of chemokines using the GSE5081 dataset. Supplementary fig. 3 Chemokine CCL3 distribution in cell types and gastric tissue. A-B Analysis the distribution of the chemokine CCL3 in cell types and gastric tissue using THE HUMAN PROTEIN ATLAS database (https://www.proteinatlas.org/). Supplementary fig. 4 The verification of CCL3 overexpression efficiency. A-B The RNA and protein expression level of CCL3 after transfected CCL3 overexpressed-plasmid in THP-1 cells. C The level of CCL3 in the medium of CCL3 overexpressing macrophages. Abbreviations: PV, pcDNA vector. The data are presented as the mean±S.D. after triplicate. Two groups were compared by t-test. Note: *P &lt; 0.05. Supplementary fig. 5 STAT1 phosphorylation level in mice gastric mucosa. A The protein expression level of P-STAT1 in the gastric mucosa of mice infected or uninfected with H. pylori (PBS: n=2; H. pylori: n=3). Supplementary fig. 6 Exploration of pathways for gastric mucosal barrier damage caused by conditioned media and CCL3. A The phosphorylation level of P65 in GES-1 and MKN28 cells treated with different culture media. B The phosphorylation level of P65 in GES-1 treated with recombinant CCL3 protein (160 ng/ml). C The phosphorylation level of P65 in GES-1 and MKN28 cells treated with CCL3 overexpressing macrophage medium. Abbreviations: PV, pcDNA vector. Supplementary fig. 7 The phosphorylation level of P38 in mice injected with CCL3 recombinant protein. A IHC detected the phosphorylation level of P38 in mice gastric mucosal (NC: n=4; CCL3: n=6). Abbreviations: IHC, immunohistochemical. Two groups were compared by t-test. Note: **P &lt; 0.01.