Fig. 6

AU1 scFv restrict the growth of the pdmH1N12009 virus. A549 cells were infected with pdmH1N12009 at MOI 1.0 and fed replete medium supplemented with different scFv (100 µg/ml). NP vRNA was quantified after 8 h by RT PCR analysis. The values were normalized with GAPDH mRNA levels. A NP transcript level was measured in the supernatant of the A549 cell culture. Only scFv AU1 shows the restricted release of viruses. B The intracellular level of the NP gene is not affected significantly in infected cells by any of the scFvs. C scFv AU1 shows inhibition in a dose-dependent manner. NP vRNA level was significantly reduced at 25 µg/ml. D scFv AU1 did not show a significant reduction of NP level intracellularly. E Cytopathogenic effect of influenza virus in the presence of scFv AU1 was observed. In the mock, cells were healthy. In the presence of viral infection with H1N1 in A549, it showed significant cells death. In the presence of scFv AU1 with H1N1 virus infection in A549 cells, it showed the significant decreases of cells death. The morphological changes of A549 cells at 48 h post infection under a phase contrast inverted microscope were shown. Scale bars, 100 μm. F Immunofluorescence assay showed the higher viral load in infected cells in the presence of AU1 scFv compared to mock. The viral load is represented by the M1 protein (green). The DAPI blue stain represents the nucleus of a cell. Scale bars, 15 μm. Graph showing the quantification of fluorescence intensity. (N = 40) “N” indicates the number of cells analyzed. All experiments were performed in triplicate. Statistical analysis was performed using the one-way analysis of variance (ANOVA) with Bonferroni’s correction for multiple comparisons and students’ t-test. *** P < 0.001; ** P < 0.01; * P < 0.05. Data points represent averages of triplicates, and error bars represent standard deviations