Figure 2

Activation of mTOR inhibits PPV-induced autophagy and suppresses PPV replication. PTCs were transfected with the mTOR activator Rheb plasmid or the same quality of vector and then infected with PPV for 12 h. A The levels of mTOR, p-mTOR, Beclin1 and LC3 II were examined by Western blotting. B–D Ratios of p-mTOR to mTOR, Beclin1 and LC3 II to β-actin as reported in A were calculated and analysed. E, F The mRNA levels of beclin1 and lc3 were detected by qPCR. G, H GFP-LC3-transfected PTCs were pre-treated with Rheb or DMSO. After infection with PPV or mock for 12 h, GFP-LC3 puncta formation in the cells was observed under laser scanning confocal microscopy (G). DAPI (blue) was used to stain nuclear DNA; scale bar: 15 μm. H The number of GFP-LC3 puncta in each cell was counted to calculate the average number of GFP-LC3 puncta per cell. At least 50 cells were counted in each group. I PTCs were infected with PPV after treatment or mock treatment with Rheb, and PPV DNA copies were then evaluated by Q-PCR 24 h post-infection. The results are the mean ± SEM (SD) of three experiments, * p < 0.05; ** p < 0.01.