Figure 6: Pharmacological inhibition of autophagy enhances TGEV replication.

(a,d) ST cells were treated with wortmannin (3 μM), LY294002 (25 μM) or DMSO carrier control for 4 h prior to TGEV infection. At 24 h post infection, cells were lysed and subjected to western blot with the antibodies against LC3 or β-actin. (b,e) Immunofluorescence assay to detect TGEV with mAb 3D7. Cells were treated with control or autophagy inhibitors for 4 h indicated, followed by infection with TGEV H165 at MOI of 1. At 24 h after infection, cell monolayer was stained for TGEV N protein expression and nuclei counterstained. The percentage of TGEV-infected cells per view from three independent experiments is expressed as mean ± SD. (c,f) TGEV titers on ST cells treated with wortmannin, LY294002, or control. The progeny virus titers were calculated and expressed as TCID₅₀ per 0.1 ml. Results represent the mean ± SD for three independent experiments. *p < 0.05. The p value is calculated using Student’s t-test. Gels were run under the same experimental conditions. For better clarity and conciseness of the presentation, cropped blots are shown. The raw uncropped images can be found in the Supplementary Fig. S4.