Figure 4

CagA-dependent repression of primary (pri-)miR-371-372-373 and upregulation of large tumor suppressor homolog 2 (LATS2) expression . (A,B) Expression of the primary miR-371-372-373 cluster transcript (A) or LATS2 mRNA (B) was determined by quantitative real time PCR (RT-qPCR) in uninfected AGS cells or cells infected for 24 h at multiplicity of infection (MOI) 100 with either Helicobacter pylori 26695 or its ΔCagA or ΔCagE isogenic mutants. Bars indicate the relative pri-micro RNA (miRNA) or LATS2 mRNA expression, normalized to the ribosomal protein P0 mRNA and compared to uninfected AGS cells. Data are mean ± SD of three independent experiments. **P < 0.01; ***P < 0.001. (C) LATS2 immunoblot of AGS cells infected or not with H. pylori or its isogenic mutants as described above. α-Tubulin was used as a loading control. (D) Enhanced green fluorescent protein (EGFP) fluorescence of LATS2 reporter cells infected or not with wild-type H. pylori, or the ΔCagA or ΔCagE isogenic mutants. Cells were infected as described above and observed 24 h later on an inverted microscope (Zeiss) in phase contrast (right panels) or epifluorescence (left panels).