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. 2020 Mar 31;11(2):e03350-19. doi: 10.1128/mBio.03350-19

FIG 5.

FIG 5

BspB and RicA modulate Golgi apparatus-associated vesicular traffic in a Rab2a-dependent manner. (A) Representative confocal micrographs of GS15 steady-state distribution in Brucella-infected BMMs transduced to express GFP-GS15, showing either Golgi apparatus-localized (normal) or diffuse patterns. Golgi structures were labeled using an anti-GM130 antibody. Scale bar, 10 μm. (B) Representative Western blotting of Rab2a depletion in BMMs. BMMs were nucleofected with either nontargeting (siNT) or siRab2a siRNAs, and Rab2 levels were evaluated after 72 h via Western blotting of Rab2a and β-actin as a control. (C) Quantification of GFP-GS15 redistribution from the Golgi apparatus in BMMs treated with either nontargeting (siNT) or siRab2a siRNAs and then left uninfected (control) or infected for 24 h with wild-type (2308), ΔbspB, ΔricA, ΔbspB ΔricA, or ΔbspB ΔricA::ricA B. abortus strains. Values are means ± SD of results from 3 independent experiments. Asterisks indicate a statistically significant difference between tested conditions, assessed using one-way analysis of variance (ANOVA) followed by a Dunnett’s multicomparison test. ns, not significant. (D) Representative confocal micrographs of BMMs cotransduced to express GFP-GS15 with mCherry (control), HA-BspB, or myc-RicA, showing normal or diffuse distributions of GFP-GS15. Scale bar, 10 μm. (E) Quantification of GS15 redistribution in BMMs cotransduced to express GFP-GS15 with mCherry, HA-BspB, myc-RicA, or HA-BspB and myc-RicA. Values are means ± SD of results from 3 independent experiments. Asterisks indicate a statistically significant difference compared to control cells, assessed using one-way analysis of variance (ANOVA) followed by a Dunnett’s multicomparison test. ns, not significant.