VE-2B interacted directly with SAP102, and VE-2B, SAP102 and PSD-95 formed co-clusters.

<p>(A) Western blotting was performed on cortical neurons infected with VE-2B and immunoprecipitated with i1 antibody to VSVG at 10 minutes and 3 hours after switch to permissive temperature (24 fold enrichment). Immunoblots were performed with rabbit anti-VSVG, PSD-95, and SAP102. Input, unbound, and immunoprecipitation fractions (IP) were run for each blot. To minimize the possibility of false-positives, buffer only was added to the lanes adjacent to the VE-2B IP lanes. Identical film exposure times for both anti-PSD-95 and anti-SAP102 indicate that SAP102 associates at 10 minutes and 3 hours after ER exit whereas PSD-95 does not. Longer film exposure times did not unambiguously indicate that PSD-95 was directly associated. (B) Western blotting was performed on cortical neurons infected with VE and immunoprecipitated with antibody to VSVG at 10 minutes after switch to permissive temperature. Immunoblots were then performed with rabbit anti-VSVG, and SAP102. Input, unbound, and IP fractions were run for each blot. The results indicated that VE (left panel) did not co-immunoprecipitate measurable quantities of SAP102 (right panel). (C) VE-2B transfected neurons 3 hours after ER release were immunostained for PSD-95 (red) and SAP102 (blue; scale bar, 5 µm). Aqua-colored arrows indicate VE-2B co-localized with SAP102 primarily. White arrows indicate VE-2B co-localized with both PSD-95 and SAP102.