Fig. 2: Dynein plays a role in moving BCV membrane remnants during Shigella infection.
A Time-lapse microscopic images of the recruitment of dynein, as marked by DYNIC2 (gray) in transfected HeLa cells. Galectin-3-mOrange (red) was used as a marker of vacuolar rupture. Images were captured every min and maximum z-projection of stacks of the representative infection focus is shown. Blue arrowheads indicate the localization of dynein on BCV remnant while the magenta arrowheads indicate some dynein-positive infection-associated macropinosomes. Intracellular bacteria were indicated by white dotted ovals. Scale bars are 5 μm. B Analysis of the fates of individual Shigella in control HeLa cells and cells subjected to p150Glued subunit depletion (siRNA p150Glued). The bars represent mean ± SEM of three independent replicates. Time-lapse microscopic analyses of Shigella infection of cells subjected to RNA interference of non-targeting control (siRNA Neg) versus p150Glued subunit depletion (siRNA p150Glued) were performed to examine C time of Shigella BCV disassembly and D Shigella escape time (i.e. formation of actin tails). n > 90 infected cells in three independent replicates in each condition. The bars (magenta) represent the mean and unpaired t-tests were performed. E Analysis of the recruitment of LC3 to intracellular Shigella in infected cells at 45 min-post infection in cells subjected to RNA interference of non-targeting control (siRNA Neg) versus p150Glued subunit depletion (siRNA p150Glued). n > 85 infected cells in three independent experiments in each condition. Data are represented by mean ± SEM of three independent replicates. Statistical analysis used two-tailed Welch’s t-test, with reported p-values for significance comparison (ns non-significant; ****p < 0.0001). Source data are provided as a Source Data file.
