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. 2017 Feb 8;7:29. doi: 10.3389/fcimb.2017.00029

Figure 4.

Figure 4

The non-structural protein 3D and 3C of EV-D68 mediates cell cycle alterations. (A,C) Effect of 3D and 3C on cell cycle progression. Left panel: The distribution of cell cycle in 293T cells was analyzed at 36 h after transfection with 2 μg of VR1012-3D-HA (3D), VR1012-3C-HA (3C) or the corresponding control vector VR1012 (mock). Right panel: The ratio of cells in G0/G1 to G2/M at 36 h after transfection with 2 μg of VR1012-3D-HA (3D), VR1012-3C-HA (3C) or the corresponding control vector VR1012 (mock). (B,D) The cell cycle distribution in 293T cells was analyzed at 36 h after transfection with 0, 0.5, 1, or 2 μg plasmid as indicated. Left panel: The histograms show the percentage of each phase in the cell cycle. Right panel: The ratios of cells in G0/G1 to G2/M. (E,F) The expression of cell cycle-related proteins after transfection of 293T cells with 3D, 3C or corresponding vector (mock) was assessed by Western blot analysis at 36 h. Histone is shown as a loading control. (G) Model for the distinct effects of the non-structural proteins 3D and 3C in cell cycle arrest caused by EV-D68. The results indicate the mean ± S.D of three independent experiments. **P < 0.01 and ***P < 0.001.