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. 2019 Oct 24;9(2):257–276. doi: 10.1016/j.jcmgh.2019.10.007

Figure 5.

Figure 5

LATS2 and YAP1 regulate each other both in basal conditions and in response to H pylori infection. (A) Western blot analysis of LATS2, LATS2-PThr1041, YAP1, and YAP-PSer127 protein levels in AGS, MKN74, and RPE1 cells infected or not with the 7.13 WT H pylori (24 hours of HPI) 30 hours after the last transfection with small interfering RNAs, either negative control (siCtrl), or against LATS2 (siLATS2-1 and siLATS2-2) or YAP1 (siYAP1-1 and YAP1-2). The numbers under each band correspond to the quantification of the relative protein expression normalized to that of the housekeeping protein glyceraldehyde-3-phosphate dehydrogenase (GAPDH) or α-tubulin. (B) Comparison of LATS2 and YAP1 basal expression determined by Western blot in AGS, MKN74, NCI-87, and Kato III human gastric epithelial cell lines. (C) Relative mRNA levels of YAP1 and LATS2 expression in siRNA transfected cells at 24 hours of HPI with H pylori 7.13 WT vs noninfected cells (ratio of mRNA levels in H pylori–infected cells vs noninfected cells). (D) TEAD-luciferase reporter activity in noninfected AGS and MKN74 cells at 30 hours after transfection with siCtrl, siLATS2, siYAP, pEGFP (control), pLATS2, pYAP1, and pYAP-S127A. (E) Quantification of the TEAD-luciferase reporter activity measured at 2 and 24 hours of HPI corresponding, respectively, to 8 and 30 hours after the last siRNA transfection in AGS and MKN74 cells. (F) Relative mRNA levels of CTGF and CYR61 expression in siRNA-transfected cells at 24 hours of HPI with WT H pylori vs noninfected cells (ratio of mRNA levels in H pylori–infected cells vs noninfected cells). (C and F) The horizontal dotted lines represent the level of the noninfected siControl cells. Bars represent the mean fold changes ± SEM, 3 < n < 6. *P < .05, **P < .01, ***P < .001, and ∗∗∗∗ P < .0001 vs siControl infected/noninfected cells using the Kruskal–Wallis test. #P < .05 vs noninfected siControls using Kruskal–Wallis test. (D and E) Bars represent the mean fold changes ± SEM, 3 < n < 6. *and #P < .05, **P < .01 vs respective noninfected siControl cells using the Kruskal–Wallis test. $Unavailable data. siCtrl, siControl.