Reduced VAPA and VAPB expression does not alter intracellular bacterial burdens, and overexpressed full-length VAP proteins localize throughout the ER. (A) Number of bacteria in cells infected with sca2::Tn mutant bacteria (28 hpi) after the indicated siRNA treatment. The mean ± SEM of three independent experiments (large outlined circles) are superimposed over the raw data from individual cells (small unlined circles) (>25 cells counted per condition per independent experiment). (B) Number of bacteria in WT or VAPA/B DKO HeLa cells infected with sca2::Tn mutant bacteria (28 hpi). The mean ± SEM of three independent experiments (large outlined circles) is superimposed over the raw data from individual cells (small unlined circles) (>25 cells counted per condition per independent experiment). (C and D) Live-cell imaging snapshots at 28 hpi of sca2::Tn mutant (magenta) infecting A549-mCherry-Sec61β cells (cyan) transiently expressing HaloTag-VAPA (A) or HaloTag-VAPB (yellow) (B). Scale bar, 5 µm; inset, 2.5 µm.