Figure 5

Analysis of ORF2p generated from a functional wild-type and a functional codon-optimized full-length L1 expression plasmids in 293 cells. (Top) Western blot analysis of ORF2p generated from expression plasmids containing a full-length wild-type L1 (L1wt), a full-length codon-optimized L1 (L1co), or a codon-optimized ORF2 transiently transfected in 293 cells with supernatant collected from cultured hybridoma cells producing anti-ORF2 antibody; 293 cells were transfected with 1, 2, or 4 μg of the L1wt or L1co expression plasmids or 2 μg of the ORF2 expression plasmid and total protein was harvested 24 hours after transfection. Control lane indicates cells transiently transfected with an empty vector. Positions of molecular markers are indicated on the right as 100 or 150 kDa. (Bottom) The same experiment and analysis as in (top), but using secondary antibodies only. Positions of molecular markers are indicated on the right as 100 or 150 kDa. Total amount of 293 cell lysate loaded is in μg.