Release of Intracellular Calcium Stores Facilitates Coxsackievirus Entry into Polarized Endothelial Cells
Figure 1
The CVB entry mechanism is distinct between polarized endothelia and epithelia.
(A) Dynasore (100 µM), dynaminK44A, and dynamin II siRNA all significantly inhibit CVB infection in HBMEC and HAEC, but have no effect on CVB infection in Caco-2 cells. Data are normalized to DMSO control, wild-type dynamin II, or control siRNA-infected cells. (B) Immunofluorescence-based assay for viral internalization in HBMEC and Caco-2 cells pre-treated with DMSO (control) or dynasore and exposed to CVB (MOI = 50) for 1 hr at 37°C. Red fluorescence (or overlapping red and green fluorescence) indicates virus on the cell surface; green fluorescence in the absence of red indicates internalized virus. (C) HBMEC monolayers expressing dominant-negative or wild-type forms of caveolin-1, caveolin-3, or Eps15 were exposed to CVB (MOI = 1) and stained for VP1 at 14 hr p.i. The graph shows the number of transfected cells expressing VP1 as a percent of control infections (dashed line). (D) HBMEC monolayers exposed to CVB (MOI = 50) and Alexa-Fluor cholera toxin-B (CTB – red) were stained for caveolin-1 (blue) and VP1 (green) at 60 min p.i.