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. 2018 Feb 20;9(1):e01800-17. doi: 10.1128/mBio.01800-17

FIG 5 .

FIG 5 

Y. pestis recruits Rab4a and Rab11b to the YCV. (A and B) Representative confocal microscopy images of primary peritoneal macrophages infected with Y. pestis CO92 pCD1(-) pGEN::mCherry (Live) (MOI of 3), PFA-killed Y. pestis CO92 pCD1(-) pGen::mCherry (Killed) (MOI of 3), or E. coli K-12 pGEN::mCherry (E. coli) (MOI of 20) and labeled with anti-Rab4a (A) or anti-Rab11b (B) antibodies. Merged (bacteria [red] and Rab protein [green]) and YCV-Rab colocalization fields generated by Imaris (COLOC) are shown. Bars = 5 µm. (C) Frequency of colocalization of bacterium-containing vacuoles with endogenous Rab4a in peritoneal macrophages. (D) Frequency of colocalization of bacterium-containing vacuoles with Rab4a-EGFP in RAW264.7 macrophages transfected with pEGFP-Rab4a. Yp, Y. pestis; Ec, E. coli. (E) Frequency of colocalization of bacterium-containing vacuoles with endogenous Rab11b in peritoneal macrophages. (F) Frequency of colocalization of bacterium-containing vacuoles with Rab11b-EGFP in RAW264.7 macrophages transfected with pEGFP-Rab11b. (G) Frequency of YCV colocalization with Rab4a-EGFP during 10 h of Y. pestis CO92 pCD1(-) pGEN::mCherry infection of RAW264.7 macrophages transfected with pEGFP-Rab4a. (H) Frequency of YCV colocalization with Rab11b-EGFP during 10 h of Y. pestis CO92 pCD1(-) pGEN::mCherry infection of RAW264.7 macrophages transfected with pEGFP-Rab11b. (I and J) Representative images of RAW264.7 macrophages transiently transfected with pEGFP-Rab4a (I) or pEGFP-Rab11b (J) and coinfected with Y. pestis CO92 pCD1(-) (blue) (MOI of 3) or E. coli K-12 pGEN::mCherry (red) (MOI of 20). Bars = 10 µm. (K and L) Frequency of colocalization of Y. pestis- or E. coli-containing vacuoles in coinfected cells withEGFP-Rab4a (K) or EGFP-Rab11b (L). One-way ANOVA with Tukey’s posthoc test was performed, and the results are indicated as follows: ns, not significant; *, P ≤ 0.05; **, P ≤ 0.01; ***, P ≤ 0.001.