Fig 3.
POWV NS1 mutant dimerization and glycosidase analysis. (A) VeroE6 cells were infected with WT POWV LI9, recLI9-NS1N85Q, recLI9-NS1N208Q, or recLI9-NS1N224Q mutants (MOI, 1). Cell lysates were harvested 7 dpi with or without heat-directed dimer disassembly prior to Western blot analysis using anti-POWV-NS1 mAb (1:5,000) or anti-GAPDH (1:5,000). (B) VeroE6 cells were infected with WT LI9, recLI9-NS1N85Q, recLI9-NS1N208Q, or recLI9-NS1N224Q mutant viruses (MOI, 1). Cell lysates were harvested 7 dpi, and 20 µg of protein lysate or 50 µL of supernatant was subjected to Endo H or PNGase F digestion. Undigested and digested samples from cell lysate or supernatant were analyzed by Western blot using antibody to POWV-NS1 or GAPDH.