Figure 8.

Cholesterol depletion does not affect HGF-R recruitment or initial membrane recruitment at the InlB entry site but subsequent closure of the phagocytic cup. (A) Control and cholesterol-depleted Vero cells were incubated for 30 min with InlB-coated beads. Cells were washed three times, fixed, permeabilized, and labeled for HGF-R (clone DL21; Alexa Fluor 546–conjugated secondary antibodies) and for F-actin (Alexa Fluor 488 phalloidin). Phase-contrast and fluorescent images were acquired with a 100× objective. Arrows indicate the InlB-coated beads that recruited HGF-R. Arrowheads indicate the InlB-coated beads that recruited HGF-R and F-actin. Bar, 10 μm. (B) Percentage of InlB-coated beads which have recruited HGF-R. (C) Control and cholesterol-depleted Vero cells were incubated for 5, 15, and 30 min with InlB-coated beads. Cells were washed three times and processed for scanning EM as described in Materials and methods. Extracellular portion of the beads were colorized using the Adobe photoshop software. Bar, 1 μm.