Journal: Cell communication and signaling : CCS

Article Title: Helicobacter pylori disrupts gastric mucosal homeostasis by stimulating macrophages to secrete CCL3.

doi: 10.1186/s12964-024-01627-5

Figure Lengend Snippet: Fig. 3 H. pylori infection promotes the expression of CCL3. A Analysis chemokines in the medium of THP-1 co-cultured with H. pylori. B Differential analysis of chemokines about the GSE27411 dataset. C, D The protein and RNA expression level of CCL3 in THP-1 co-cultured with H. pylori. E The secretion level of CCL3 in the medium of H. pylori -infected macrophages. F The protein expression level of CCL3 in H. pylori infected mice gastric tissue (PBS: n = 2; H. pylori: n = 3). G The RNA expression level of CCL3 in H. pylori infected mice gastric tissue (PBS: n = 3; H. pylori: n = 5). H IHC analysis CCL3 expression in mice (PBS: n = 3; H. pylori: n = 6). I IHC analysis CCL3 expression in human (each group: n = 4). J The expression level of CCL3 in mice serum (each group: n = 4). K The expression level of CCL3 in human serum (each group: n = 3). Abbreviations: IHC, immunohistochemical. 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 < 0.05, **P < 0.01, ***P < 0.001

Article Snippet: Human CCL3 ELISA kit was purchased from CUSABIO (CSB-E04662h, China).

Techniques: Infection, Expressing, Cell Culture, RNA Expression, Immunohistochemical staining

Journal: Cell communication and signaling : CCS

Article Title: Helicobacter pylori disrupts gastric mucosal homeostasis by stimulating macrophages to secrete CCL3.

doi: 10.1186/s12964-024-01627-5

Figure Lengend Snippet: Fig. 5 CCL3 neutralizing antibody and CCL3 receptor inhibitor improve mucosal damage and inflammatory response. A Effect of CCL3 neutralizing antibody (100 ng/ml) on TEER of MKN28 cells. B The protein expression level of the tight junction in GES-1 cells treated with CCL3 neutralizing antibody in conditioned media. C IF detected tight junction between MKN28 cells treated with CCL3 neutralizing antibody in conditioned 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. D, E Apoptosis and proliferation of GES-1 treated with CCL3 neutralizing antibody in conditioned media (D: Annexin V- PI apoptosis flow cytometry; E: EdU). F Effect of CCL3 receptor inhibitor (Maraviroc, 100 nM) on TEER of MKN28 cells. G The protein expression level of the tight junction in GES-1 cells treated with CCL3 receptor inhibitor. H IF detected the tight junction between MKN28 cells treated with CCL3 receptor inhibitor. I, J Apoptosis and proliferation of GES-1 treated with CCL3 receptor inhibitor (I: Annexin V- PI apoptosis flow cytometry; J: EdU). Abbreviations: 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 < 0.05, **P < 0.01

Article Snippet: Human CCL3 ELISA kit was purchased from CUSABIO (CSB-E04662h, China).

Techniques: Expressing, Staining, Disruption, Flow Cytometry, Infection, Immunofluorescence

Journal: Cell communication and signaling : CCS

Article Title: Helicobacter pylori disrupts gastric mucosal homeostasis by stimulating macrophages to secrete CCL3.

doi: 10.1186/s12964-024-01627-5

Figure Lengend Snippet: Fig. 7 H. pylori-infected macrophage culture medium and chemokine CCL3 disrupt gastric mucosal barrier through P38 phosphorylation. A The P38 phosphorylation level in GES-1 treated with different conditioned media. B IF detected P38 phosphorylation levels in GES-1 treated with different conditioned media. DAPI was used for nuclear staining (blue), P-P38 stained in red. C, D The P38 phosphorylation level in GES-1 cells with recombinant CCL3 protein (160 ng/mL) and CCL3 overexpressing macrophage medium. E, F The P38 phosphorylation level in GES-1 cells treated with CCL3 neutralizing antibody (100 ng/mL) and CCL3 receptor inhibitor (Maraviroc, 100 nM). G Effects of P38 phosphorylation inhibitor (SB203580, 10 μM) on TEER of MKN28 cells. H IF detected the effect of P38 phosphorylation inhibitor on tight junctions between MKN28 cells. DAPI was used for nuclear staining (blue), Occludin was used for tight junction staining (green), arrows showed the disruption of tight junctions between cells. I, J The protein expression level of tight junction in GES-1 cells treated with P38 phosphorylation inhibitor. K, L Apoptosis and proliferation of GES-1 treated with P38 phosphorylation inhibitor (K: Annexin V- PI apoptosis flow cytometry; L: EdU). Abbreviations: NC, normal culture medium; MC, macrophage culture medium; HMC, H. pylori-infected macrophage culture medium; PV, pcDNA vector; 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 < 0.05, **P < 0.01

Article Snippet: Human CCL3 ELISA kit was purchased from CUSABIO (CSB-E04662h, China).

Techniques: Infection, Phospho-proteomics, Staining, Recombinant, Disruption, Expressing, Flow Cytometry, Plasmid Preparation, Immunofluorescence

Journal: Cell communication and signaling : CCS

Article Title: Helicobacter pylori disrupts gastric mucosal homeostasis by stimulating macrophages to secrete CCL3.

doi: 10.1186/s12964-024-01627-5

Figure Lengend Snippet: Fig. 9 Schematic diagram of the mechanism of gastric mucosal damage caused by CCL3 which is secreted by H. pylori infected macrophages. H. pylori induces the expression of the chemokine CCL3 by macrophages via the JAK1-STAT1 pathway, and CCL3 disrupts the gastric mucosal barrier via P38 phosphorylation

Article Snippet: Human CCL3 ELISA kit was purchased from CUSABIO (CSB-E04662h, China).

Techniques: Infection, Expressing, Phospho-proteomics