Fig. 2. Expression of complex-glycosylated and basal-glycosylated recombinant hemagglutinin in lettuce. (A) Schematic representation of portion of expression cassette of pBYR11ek2Md flanking the cloned canine HA (H3N2) with N-SP at N-terminal (upper) or KEDL at C-terminal (lower). (B, C) Immunodetection of complex-glycosylated and basal-glycosylated rHA in leaves agroinfiltrated with HA in geminiviral vector into tobacco (B) and lettuce (C). (D) Predicted N-glycosylation site (N-Glyc) in HA sequence. N-Glyc results column output “+” represents potential > 0.5 and output “++” represents the potential > 0.5 and the jury agreement (9/9) where the potential score is the average of the output of the nine neural network and default threshold is 0.5. Western blot analyses were performed using polyclonal anti-H3N2 HA antibody. ‘Housekeeping protein’ RuBisCo, detected by Coomassie staining analysis, is shown as a loading control. N-SP, native signal peptide; rHA, recombinant hemagglutinin; HA, hemagglutinin; UTR, untranslated region; LIR, long intergenic region; SIR, short intergenic region; 35S, cauliflower mosaic virus (CaMV) 35S promoter; 3′ MAR, 3′ elements of tobacco Rb7 chromatin matrix attachment region.