FIG. 6.

Comparison of saponin-depleted and nondepleted cell extracts during poliovirus infection and 2BC expression. (A) COS-7 cells were transfected with GFP-LC3 expression plasmid and then 48 h later infected with poliovirus (50 PFU/cell). Cells were then harvested at 0, 2, 4, and 6 h postinfection. (B) COS-7 cells were transfected with a plasmid that expressed GFP-LC3 and increasing proportions of 2BC-expressing plasmids relative to a control vector, followed by harvesting. Recovered cell samples were split in half and either subjected to extraction first with 0.5% saponin and then with NP-40-containing lysis buffer or directly treated with NP-40-containing lysis buffer. Cell extracts were separated by SDS-PAGE on a 10% polyacrylamide gel, transferred to PVDF, and blotted with anti-LC3 and then anti-VSV-G antibodies.