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. 2020 Mar 5;21(5):1801. doi: 10.3390/ijms21051801

Figure 2.

Figure 2

Production of IL-33 in cell cultures exposed to H. pylori components. (A) Representative images of guinea pig primary gastric epithelial cells or fibroblasts treated with control siRNA- labelled with fluorescein isothiocyanate; green (FITC) and photographed in the confocal microscope (Leica TCS SP Wetzlar, Germany), at magnification ×100. (B) Production of IL-33 by the guinea pig primary gastric epithelial cells not transfected or transfected with siRNA IL-33, unexposed or exposed on H. pylori glycine acid extract (GE). (i) Level of intracellular IL-33 in the guinea pig primary gastric epithelial cells measured as relative fluorescence units (RFU). (ii) Representative images of the guinea pig primary gastric epithelial cells stained for intracellular IL-33 (green) and counterstained with 4′,6-diamino-2-phenylindole (DAPI; blue) or phalloidin –Texas RedTM (red), photographed in the confocal microscope (Leica TCS SP, Wetzlar, Germany), at magnification ×63. (iii) Concentration of IL-33 in the supernatants of guinea pig primary gastric epithelial cells estimated by the enzyme-linked immunosorbent assay (ELISA). (C) Production of IL-33 by the guinea pig fibroblasts, not transfected or transfected with siRNA IL-33, unexposed or exposed on H. pylori GE. (i) Level of intracellular IL-33 in the guinea pig fibroblasts measured as relative fluorescence units (RFU). (ii) Representative images of the guinea pig fibroblasts stained for intracellular IL-33 (green) and counterstained with DAPI (blue) or phalloidin (red) and photographed in the confocal microscope (Leica TCS SP, Wetzlar, Germany) at x63 magnification. (iii) Concentration of IL-33 in supernatants of guinea pig fibroblasts estimated by the ELISA. Results are showed as median with the range of five experiments performed in triplicates for each experimental variant. Statistical analysis was performed using the nonparametric U Mann-Whitney test with significance for *, ** p < 0.05,* unstimulated vs. GE stimulated untransfected cells, ** untransfected vs. transfected cells in medium only or in medium with GE.