LATS2 restricts H pylori–induced EMT in AGS cells. (A) Phase-contrast images of cells in siCtrl- and siLATS2-transfected cells upon 24 hours of HPI with H pylori 7.13 WT in AGS and MKN74. Scale bars: 20 μm. (a) In AGS cells (left panel), Yellow arrows indicate cells with the elongated hummingbird phenotype appearing in siLATS2 cells and at 24 hours of HPI. (b) In MKN74 cells (right panel), Yellow arrows indicate loosely attached round cell clusters appearing in siLATS2 cells infected or not infected. (B) Percentage of cells with the hummingbird phenotype in AGS cells. *P < .05 vs the respective noninfected condition. ∗∗∗∗P < .0001 vs the respective noninfected condition. #### P < .0001 vs H. pylori–infected controls. $$$$ P < .0001 vs noninfected control cells in analysis of variance test. (C) Relative levels for the EMT markers ZEB1 (dark grey bars), BMP1 (light grey bars), MMP9 (crosshatched bars) mRNAs, measured at 24 hours of HPI with H pylori 7.13 WT in siControl-, siLATS2-1–, and siLATS2-2–transfected AGS, MKN74, and RPE1 cells. Bars represent the mean fold changes ± SEM of RT-qPCR data, 3 < n < 6. ∗P < .05, ∗∗P < .01, ∗∗∗P < .001, ∗∗∗∗P < .0001 vs noninfected siControl using the Kruskal–Wallis test. ##P < .01 vs H pylori–infected siControl using the Kruskal–Wallis test. (D and E) Expression and subcellular localization of (D) ZEB1 and (E) MMP9 (in green) assessed by immunofluorescence in AGS, RPE1, and MKN74 cells. F-actin was marked with Alexa-546–labeled phalloidin (red) and nuclei with 4’-6-diamino-phenyl-indol (blue). Scale bars: 10 μm. (F) Invasion capacity through collagen 1–coated Transwell inserts of H pylori–infected and noninfected siControl- and siLATS2-transfected AGS, MKN74, and RPE1 cells, and of noninfected pEGFP- (control) and pLATS2-transfected AGS cells, at 24 hours of HPI. Bars represent the mean fold changes ± SEM (n = 5) of the number of cells having crossed the Transwell 18 hours after plating, compared with noninfected siControl cells. **P < .01, ***P < .001, and ****P < .0001 vs noninfected siControl using the Kruskal–Wallis test. siCtrl, siControl.