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. 2016 Feb 10;6:20839. doi: 10.1038/srep20839

Figure 6. Effect of WT or M2 mutant 5′UTR of CA16 on viral RNA synthesis and viral translational capacity in transfected HEK293T cells.

Figure 6

HEK293T cells were transfected with same amount of the WT or M2 infectious clone or empty vector, and were harvested at 12 h, 24 h, 36 h and 48 h after transfection. (A) RNA was extracted from a portion of each sample, treated with Dnase to degrade the transfected plasmid. DNA and then analyzed by RT-qPCR with primers specific for GAPDH or CA16 VP1 RNA. GAPDH was used as a control. The RNA level obtained by transfection with WT 5′UTR was normalized to 100%. (B) A portion of each cell sample was used to detect the VP1 protein by Western blot. (C) A portion of each cell sample was used to detect the viral titer. (D) Viral loads in supernatant of transfected HEK293T cells at 48h after transfection were detected by RT-qPCR with primers specific for CA16 VP1 RNA. (E) The VP1 protein of the WT or M2 virus in the supernatant was detected by Western blot using VP1 antibody. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001.