Figure 7.

Polarized expression of receptors on ECC-1 cells. (A) ECC-1 cells were grown on Transwells and stained with EZ-Link Sulfo-NHS-Biotin to detect cellular plasma membranes (red). Then, they were permeabilized, and the nuclei were detected by DAPI (blue). Syndecan, nectin-1, integrinαv, and IP₃R were visualized by labeling each with a cocktail of specific mAbs (green). Successive axial sections from the apical to the basal membrane were obtained; the axial images 1.5 and 7.5 μm from the apical surface and Z-sections are representative of at least five independent experiments. (B) Representative XYZ-section images of Calcium Green–loaded polarized ECC-1 cells after infection from either the apical (AP) or basolateral membrane (BL) with DiD-labeled HSV-2(G). Cells were fixed and stained 5 or 30 min after infection (plasma membrane, red; viral envelope, blue; calcium, green; colocalization of calcium and plasma membranes, yellow; colocalization of membrane and envelope, purple; and colocalization of envelope and calcium, turquoise). Results are representative of those obtained in three independent experiments.