FIG. 5.

Solubility of gE in the nonionic detergent TX100. HEC-1A cells were infected with both Ad(E1⁻)gE and Ad(E1⁻)gI for 48 h, washed with Tris-saline, and then scraped into extraction buffer containing 0.1% TX100. Cell extracts were centrifuged at 20,000 × g for 30 min. Proteins in the pellets (P) and supernatants (S) were denatured in buffer containing 2% SDS and 2% β-mercaptoethanol; then the proteins were boiled, subjected to electrophoresis, and transferred to nitrocellulose. gE was detected by incubating the blots with MAb II-481, and E-cadherin was detected with an anti-E-cadherin MAb; in each case, this was followed by incubation with horseradish peroxidase-coupled anti-mouse IgG antibodies and ECL (Amersham).