Fig. 6.

Interference with host endocytic and retromer-mediated trafficking pathways allows partial rescue of vacuole integrity defect. (A) Effect of si-Rab5B on vacuole integrity. (A, Left) immunoblot of SDS-polyacrylamide gel probed with anti-Rab5B after depletion with si-Rab5B or control scrambled si-RNA. (A, Right) the percentage of cytosolic bacteria from BMDMs treated with si-Control or si-Rab5B, and challenged with noted L. pneumophila strains for 2 h, followed by probing as described (Materials and Methods). (B) Effect of si-SNX1 on vacuole integrity. (B, Left) immunoblot of SDS-polyacrylamide gel probed with anti-SNX1 after depletion with si-SNX1 or control scrambled si-RNA. (B, Right) the percentage of cytosolic bacteria from BMDMs treated with si-Control or si-SNX1 and challenged with noted L. pneumophila strains for 2 h, followed by probing as described (Materials and Methods). For immunoblotting, β-actin was used as loading control. The data shown are mean ± SEM, using three biological replicates, with at least 100 LCVs counted per biological replicate. Statistical analysis was conducted using unpaired two-tailed Student’s t test and significance displayed as **P < 0.01 or ****P < 0.0001.