Figure 4.
Far western blot analysis to show rfhSP-D binding to purified (A) pH1N1 and (B) H3N2: 10 μl of concentrated virus (1.36 × 106 pfu/ml) was first run on the SDS-PAGE under reducing conditions, and then transferred onto a nitrocellulose membrane and incubated with 5 µg of rfhSP-D. The membrane was probed with anti-rabbit SP-D polyclonal antibodies. rfhSP-D bound to HA (70 kDa) and M1 (27 kDa) in the case of both pH1N1 and H3N2 subtypes. (C) ELISA to show the binding of rfhSP-D to purified recombinant hemagglutinin (HA) (μg/ml). VSV-G was used as a negative control. The data were expressed as mean of three independent experiments carried out in triplicates ± SEM. Significance was determined using the unpaired one-way ANOVA test (***p < 0.0001) (n = 3).