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. 2019 Aug 15;15(8):e1007892. doi: 10.1371/journal.ppat.1007892

Fig 4. Ratio of M1 to M2 protein expression is high in DF-1 Cells irrespective of viral M segment host origin.

Fig 4

DF-1 cells were inoculated at a MOI of 5 PFU/cell, with PR8 viruses encoding avian or human-derived M segments, and incubated at 37°C for 8 h, then lysed. Western immunoblot analysis of virus-infected DF-1 cells: (A) Vinculin expression was measured to allow normalization of viral protein levels. NP expression was measured to assess viral replication. Levels of M1 and M2 protein expression were assessed using an antibody (Mab E10) to a common epitope at the amino terminus of M1 and M2 proteins, allowing relative expression to be assessed. Levels of LC3B I and II were assessed using an antibody that detects both the precursor and activated forms of LC3B protein. (B) M1 protein and (C) M2 protein were normalized to vinculin, quantitated and displayed as a percentage of total protein expressed from the M gene. (D) The ratio of M1:M2 protein expression. (E) LC3B I protein and (F) LC3B II protein were normalized, quantitated and displayed as a percentage of total LC3B protein. Graphs in B-F show the means with SD from three independent experiments. For each experiment, two replicate Western immunoblots were performed and quantitated. Statistical significance was assessed using ordinary one-way ANOVA.