Effect of depletion of ezrin on bacterial uptake by TG cells. (A) TG cells were treated for 48h with siRNA targeting ezrin or without it (Cont.), radixin, moesin, or the negative control siRNA (QIAGEN AllStars Negative Control) (NC). Depletion of ezrin was examined by immunoblotting. Radixin was used as a comparable control. (B) Distribution of ezrin in non-treated (Control) and ezrin-depleted (siRNA ezrin) TG cells. Scale bar indicates 50 μm. (C) Bacterial uptake by TG cells. Killed or live B. abortus were deposited onto ezrin-depleted TG cells. TG cells were also treated with IFN-γ at 1000 U/ml for 12 h before inoculation of heat-killed bacteria. This result of ezrin depletion corresponds to panels A and B. Data are the averages of triplicate samples from three identical experiments. Error bars represent standard deviation. Statistically significant differences from control are indicated by asterisks (*, P < 0.01 and **, P < 0.05). (D) Distribution of ezrin (red) and heat-killed B. abortus (green) in non-treated (Control) and ezrin-depleted (siRNA ezrin) TG cells after 30 min of bacterial inoculation. TG cells were also treated with IFN-γ at 1000 U/ml for 12 h before inoculation of bacteria. Arrows point to co-localized bacteria. Scale bar indicates 10 μm.