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A second wave of Salmonella T3SS1 activity prolongs the lifespan of infected epithelial cells

Fig 4

Expression of SopB in cytosolic bacteria is sufficient to induce post-invasion Akt phosphorylation.

HeLa cells were infected with ΔsopB Salmonella complemented with plasmid borne sopB under the control of a promoter that is induced in the presence of glucose-6-phospate (PuhpT), or the arabinose inducible reporter (PBAD). (A) Induction of SopB expression in cytosolic bacteria stimulates Akt phosphorylation. Cells were fixed, and processed for PLA of phospho-Akt (pAkt) and immunostaining of SopB-2xHA (αHA). DNA was stained with DAPI. Representative confocal images are shown. Scale bars: 10 μm. (B, C) Western blot for total Akt and phosphorylated Akt (pAkt), and band densitometry, normalized to uninfected, for phosphorylated Akt, Densitometry data show means ± SD for 3 independent experiments. Western blot is from one representative experiment. Statistical significance was analyzed by 2-way ANOVA with Bonferroni’s post-test and compared to corresponding time points of ΔsopB (B) or WT (C) infected cells. * P ≤ 0.05, *** P ≤ 0.001, ns = not-significant.

Fig 4

doi: https://doi.org/10.1371/journal.ppat.1006354.g004