FIG 4.

The endosomal transporter AmtA is a component of LCVs. (A) IFC images of intact D. discoideum Ax3 that dually produced AmtA-mCherry and calnexin-GFP (pAW16) and that was infected (MOI, 5) for 2 h with mPlum-producing virulent L. pneumophila JR32 or ΔicmT(pAW14). (B) Simultaneous quantification of the IFC colocalization score between calnexin-GFP and mPlum (left) and AmtA-mCherry and mPlum (right) for >1,000 intact cells per sample at the time points postinfection indicated. (C) IFC images of LCVs from homogenized D. discoideum Ax3 that dually produced AmtA-mCherry and calnexin-GFP (pAW16) and that was infected (MOI, 50) for 2 h with mPlum-producing virulent L. pneumophila JR32 or ΔicmT(pAW14). (D) Quantification of calnexin-GFP intensity on AmtA-positive LCVs by IFC for >1,000 LCVs per sample at the time points postinfection indicated. (E) Quantification of the percentage of LCVs positive for calnexin-GFP, based on the GFP intensity on the LCV together with the results of colocalization analysis of GFP and mPlum. The data show the means and 95% confidence intervals from one representative experiment out of three independent experiments (B, D) or the means and SEMs from three independent experiments (E) (***, P < 0.001 at 2 h).