Figure 8. ESCRT‐III and Galectin‐3 are recruited to the Coxiella burnetii vacuole.

1. HeLa cells stably expressing CHMP4B‐eGFP were transfected with the mCherry‐Galectin‐3 plasmid. Twenty‐four hours later, cells were infected with WT Coxiella burnetii. Live‐cell imaging started 24 h post‐infection, and several time points are indicated during the next 24 h of observation (time indicated in minutes, see Movie EV9). Arrows indicate Coxiella‐containing vacuole (DRAQ5 labeling) becoming positive for CHMP4B‐eGFP and mCherry‐Galectin‐3. Scale bars: 50 μm.
2. Bacterial viability and replication assay upon TSG101 KD. HeLa cells were treated with either siCtrl or with two different siTSG101 for 48 h. Then, cells were infected with mCherry‐Coxiella burnetii for 48 h before lysis and serial dilutions were made to infect Vero cells. Seventy‐two hours later, infected cells were fixed, DAPI stained, and processed for quantitative image analysis. Between 30 and 40 fields representing more than 9,000 cells per condition of three independent experiments were analyzed. Error bars represent SD. Statistical significance was determined using one‐way ANOVA test. **P ≤ 0.01 and ***P ≤ 0.001.