pH neutralization causes PtdIns(3)P to persists in phagosomes. (a) RAW cells expressing 2FYVE-GFP (rainbow) were treated with 0.1% DMSO (vehicle), 1 µM ConA, 10 mM NH₄Cl, or both ConA and NH₄Cl for 15 min before the phagocytosis of IgG-opsonized beads. White and magenta arrows depict examples of 2FYVE-positive and -negative phagosomes, respectively. (b) The phagosome/cytosol ratio of 2FYVE-GFP fluorescence for each time point indicated, normalized to 7 min, represented as relative units (R.U). Data shown are means ± SEMs from three independent experiments (n = 30 for each). ANOVA test was used to compare each treatment condition to vehicle. For each time point; *, P < 0.05; **, P < 0.01. Bars, 5 µm.