Fig. 7: V-ATPase mutants show significant loss of fitness in macrophages and mice and reduced growth rates under low external pH conditions.
A Barcode trajectories of barcoded parental controls and V-ATPase knockout mutants in macrophages. B Barcode trajectories in mice. C Growth of promastigote form V-ATPase deletion mutants V1 E and V1 H and the parental cell line in standard M199 medium at pH 7.4. Cells were diluted into fresh medium to a density of 1 × 106 cells/ml every 24 h. Each point represents the average cell density measured from three separate cultures. D Growth profiles as in C, in M199 medium adjusted to pH 5.5. E Fluorescence micrographs showing the localisation of mNeonGreen-tagged V-ATPase subunits V1 G and V1 H in promastigotes (PRO) and axenic amastigotes (AXA). For each subunit, the left panel shows the merged images from phase contrast illumination, and fluorescent signals from Hoechst-stained DNA (magenta) and the mNeonGreen fusion protein (green). The right panel shows the signal from the fusion protein only. The scale bar is 10 µm. The micrographs are representative of the mNG localisation results from two independent experiments tagging these genes. Source Data for A and B are in Supplementary Data 4. Source Data for C–E are provided as a Source Data file.
