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. Author manuscript; available in PMC: 2016 May 13.
Published in final edited form as: Cell Host Microbe. 2015 Apr 23;17(5):716–725. doi: 10.1016/j.chom.2015.03.014

Figure 3. The inclusion membrane protein InaC is required for F-actin assembly at the inclusion.

Figure 3

(A) F-actin assembly at inclusions (arrowhead) is absent in HeLa cells infected with M407. F-actin (grayscale) and C. t (red) were detected by indirect immunofluorescence at 30 hpi using rhodamine-phalloidin and anti-MOMP antibodies respectively. DNA (blue) was stained with Hoechst. (B–E) F-actin assembly in M407 inclusions is rescued upon expression of wild type InaC. InaC expression is restored in M407 mutants transformed with a plasmid expressing wild type inaC from its own promoter as assessed by immunoblot (C) and immunofluorescence (red, D) analysis and without affecting the distribution of another inclusion membrane protein, IncG. F-actin assembly at the inclusion (white arrowheads, (C)) is restored in M407 transformed with a plasmid encoding inaC but not an empty vector. Note that the average intensity (D) and frequency (E) of F-actin around complemented inclusions is comparable to wild type. Mean ± SEM for three independent experiments is shown. At least 50 (F) or 300 (E) inclusions were enumerated in each triplicate for each experiment. * indicates P < 0.05 by one-way ANOVA and Newman-Keuls post hoc. Scale bars represent 10μm.