Fig 1: PPRV induces two waves of autophagy via distinct and uncoupled molecular pathways. (A and B) EECs were mock-infected or infected with PPRV or UV-irradiated PPRV (MOI = 3) for 1.5 or 12 h. The cell samples were fixed and processed for electron microscopy analysis. Scale bars, 1 μm. Magnified views of the autophagosome-like vesicles are enclosed by black square frames. Scale bars, 200 nm. (C) EECs were mock-infected or infected with PPRV or UV-irradiated PPRV (MOI = 3) for 1.5 or 12 h. The cell samples were analyzed by immunoblotting with anti-LC3, anti-SQSTM1, anti-PPRV-N and anti-ACTB (loading control) antibodies. The target protein levels relative to the ACTB levels in the mock-, UV-irradiated PPRV- and PPRV-infected EECs were determined by densitometry. (D) EECs were treated with si-control, si-ATG5 or si-GOPC and infected with PPRV (MOI = 3) for 1.5 or 12 h. The cell samples were analyzed by immunoblotting with anti-LC3, anti-SQSTM1, anti-PPRV-N and anti-ACTB (loading control) antibodies. The target protein levels relative to the ACTB levels in the siRNA-transfected cells were determined by densitometry. (E) EECs were treated with chloroquine and infected with PPRV (MOI = 3) for 1.5, 3 or 6 h. The cell samples were analyzed by immunoblotting with anti-LC3, anti-SQSTM1 and anti-ACTB (loading control) antibodies. The target protein levels relative to the ACTB levels in the treated cells were determined by densitometry. The data represent the mean ± SD of three independent experiments. Two-way ANOVA; *P < 0.05; **P < 0.01; ***P < 0.001; #P > 0.05.