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Fig. 1. Viral protein VP8 alters the transepithelial electrical resistance (TER) of epithelial monolayers (MDCK) in a reversible and dose dependent manner. (A) TER was determined in MDCK monolayers receiving 4 µg/ml of GST-VP5 (open squares) or GST-VP8 (closed triangles). In all the panels of this figure the closed squares correspond to the TER values obtained with MDCK monolayers treated with 4 µg/ml of GST. Monolayers that had been cultured for 30 minutes with 4 µg/ml of GST-VP8 (closed triangles) were washed (arrow) and transferred to media without GST-VP8 (open triangles). (B) MDCK monolayers incubated with 0.4 µg/ml (open circles), 4 µg/ml (closed triangles) or 10 µg/ml (open triangles) of GST-VP8. (C) At different times (1-5 days) after plating at confluence, the epithelial monolayers were treated with control media (gray bars) or 4 µg/ml of GST-VP8 added for 30 minutes (black bars) and 2 hours (white bars). Statistical significance was determined using a one-way ANOVA test. *P<0.05; **P<0.001; ***P<0.0001. (D) Confluent MDCK monolayers cultured for 20 hours in LC (1-5 µM Ca²⁺) were transferred to NC (1.8 mM Ca²⁺) (closed squares). Some monolayers were instead transferred to NC media containing either 4 µg/ml of VP5 (open squares) or of VP8 (closed triangles). In all these Figures, the values of TER reported as media±s.e.m. were normalized as explained in the Materials and Methods section. At each experimental point the number of monolayers on which the TER was measured is indicated.

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