Figure 9.

HSV triggers membrane Ca²⁺ release if syndecan and nectin are present, but requires integrinαv and IP₃R for the global Ca²⁺ response. (A) CaSki cell membranes were labeled with VYBRANT-DiI (red) and transfected with nonspecific or specific siRNAs as indicated. Forty-eight hours after transfection, the cells were loaded with Calcium Green and infected in a synchronized assay with DiD-envelope–labeled HSV-2(G) (blue). Live images were acquired at 1-s intervals pi at the indicated slice size. Representative images obtained 2 s after the temperature reached 37°C are shown. (B) CaSki cells were transfected with nonspecific or IP₃R siRNAs and 48 h later, cells were loaded with the Ca²⁺ indicator dye Fura-2, exposed to HSV-2(G) (moi = 10) and changes in [Ca²⁺]_i monitored. Representative results from a single experiment showing two individual cells are displayed. (C) The bar graph depicts changes in [Ca²⁺]_i elicited by HSV-2(G) on CaSki cells transfected with the indicated siRNA or CHOK1 cells. Means obtained from at least two independent experiments monitoring at least 10 cells are depicted, and the error bars indicate SE; asterisks indicate significant differences between baseline and peak Ca²⁺ response to HSV by paired t tests (p < 0.001).