FIGURE 5.

Clathrin-dependent interaction of Rab5 with BCVs during the early stage of infection. HeLa cells were pretreated with 12.5 μM CPZ and infected with B. abortus at an MOI of 10 for the indicated times. A–C, at 15 (A), 30 (B), and 60 (C) min p.i., the cells were fixed and stained with Texas red-conjugated Rab5 (red), Alexa Fluor 405-labeled B. abortus (blue), and FITC-labeled clathrin (green) antibodies. All images shown are representative of three separate experiments. Scale bars = 5 μm as indicated. D and E, the number of BCVs that colocalized with clathrin and/or Rab5 (D) or LAMP-1 (E) was determined by analyzing 100 BCVs (combined from at least five random fields) from three separate monolayers for each time point. Data represent the mean ± S.D. of triplicate trials from three independent experiments. Differences that were statistically significant compared with untreated samples are indicated. *, p < 0.05; **, p < 0.01; ***, p < 0.001.