Fig. 4

Inactivation of cagA decreased expression and nuclear localization of YAP and also downregulated transcription of YAP downstream target genes. Construction and characterization of isogenic PMSS1 ΔcagA mutants. a schematic depiction of generation of PMSS1 ΔcagA mutants. b, c Complete deletion of cagA from the genomes of the representive transformants was completely verified using primer sets F1/R1 and F2/R2 for targeting cagA (b), and Fup/54 and Fup/55 for targeting the locus containing the sequence upstream of cagA and the cat cassette (c). d Western blotting to characterize CagA expression in H. pylori cell lysates. UreB was used as a positive control. Total CagA and phosphorylated CagA in AGS cells infected with CagA⁺ H. pylori strains PMSS1 and 7.13 as well as their CagA⁻ mutant strains at MOI of 200 for 24 h were detected using Western blotting. e Cell elongation assay (Hummingbird phenotype) was performed on AGS cells infected with CagA⁺ H. pylori strains PMSS1 and 7.13 as well as their CagA⁻ mutants at MOI of 200 for 24 h. All pictures were obtained under 100× magnification. Degrees of cell elongation (lower right image) was calculated as the ratio of length to breath of a AGS cell