FIG 2.
Rab2a depletion renders BspB dispensable for rBCV biogenesis and Brucella replication. (A) Representative Western blotting of Rab2a depletion in BMMs. BMMs were nucleofected with either small interfering nontargeting (siNT) or siRab2a siRNAs, and Rab2 levels were evaluated after 72 h via Western blotting of Rab2a and β-actin as a control. (B) rBCV biogenesis in BMMs treated with either nontargeting (siNT) or siRab2a siRNAs for 72 h and infected with DsRedm-expressing wild-type (2308), ΔbspB, ΔricA, ΔbspB ΔricA, or ΔbspB ΔricA::ricA B. abortus strains. Data are means ± SD of results from 3 independent experiments. Asterisks indicate a statistically significant difference determined by two-way ANOVA with Dunnett’s multiple-comparison test. (C) Intracellular replication of DsRedm-expressing wild-type (2308), ΔbspB, ΔricA, ΔbspB ΔricA, or ΔbspB ΔricA::ricA B. abortus strains at 24 h pi in BMMs treated with either nontargeting (siNT) or siRab2a siRNAs for 72 h prior to infection. Values are means ± SD of results from at least 3 independent experiments. Asterisks indicate a statistically significant difference compared to siNT-treated BMMs infected with either wild-type (2308; green) or ΔbspB (red) strains, assessed using a nonparametric Kruskal-Wallis test with Dunn’s multicomparison statistical analysis; ns, not significant.