miR-30c Increases the Intracellular Survival of Helicobacter pylori by Inhibiting Autophagy

Figure 3

The expression of miR-30c-1-3p and miR-30c-5p is upregulated in response to H. pylori infection and downregulates xenophagy. (a) Quantitative real-time PCR detection of miR-30c-1-3p and miR-30c-5p after infection. In GES-1 cell lines, the expression of miR-30c-1-3p and miR-30c-5p rapidly increased after being infected with H. pylori (). (b) GES-1 cells were transfected with mimic (100 nM) and inhibitor (100 nM) of miR-30c-1-3p as well as miR-30c-5p, negative mimic (100 nM) or negative inhibitor (100 nM) for 36 h, afterwards infected with H. pylori for 12 h. The expressions of LC3B-II and P62 were measured by western blot. (c) Quantification of autophagosomes and autolysosomes. GES-1 cells were transfected with mimic (100 nM) and inhibitor (100 nM) of miR-30c-1-3p as well as miR-30c-5p, negative mimic (100 nM) or negative inhibitor (100 nM) for 24 h. Afterwards, mRFP-GFP-LC3 adenovirus was transfected for 24 h and finally stimulated with H. pylori for 12 h. The number of spots per cell was counted (). (d) H. pylori infected GES-1 cells for 12 h after transfection with mimic (100 nM) and inhibitor (100 nM) of miR-30c-1-3p as well as miR-30c-5p, negative mimic (100 nM) or negative inhibitor (100 nM) for 36 h. The samples were collected for TEM examination to observe ultrastructural alterations. Red arrows indicate autophagosomes, and blue arrows indicate autolysosomes. All error bars indicate SEM. , , and .

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