(A) Preparation of the recombinant HA protein. Identification of the Bacmid/HA recombinant (a) M, Marker, Lane 1, PCR product of Bacmid-HA. Recombinant HA purified from Bacmid/HA-transfected SF9 cells by Ni-NTA affinity chromatography (Coomassie Brilliant Blue staining) (b) M, prestained protein marker, Lane 1, Control (from SF9 cells transfected with blank bacmid), Lane 2, HA Purified from Bacmid/HA-transfected SF9 cells. Confirmation of HA recombinant by western blot analysis (c) M, Marker, Lane 1, HA Purified from Bacmid/HA-transfected SF9 cells, Lane 2, control. (B) Morphology changes in the recombinant HA-treated human pulmonary epithelial cells. A549 cells treated with 40 µg/ml HA (b) or the control (a) for 12 h (bar = 50 µm). The cells treated with HA become swollen, rounded and irregular in size and shape.