Figure 11: Induction of autophagy with specific siRNA targeting endogenous acetyltransferase p300 reduces TGEV replication.

(a) Autophagy is inhibited with specific siRNA targeting p300. ST cells were transfected with siRNA duplexes against porcine endogenous p300 or control siRNA at the concentration of 100 nM. Detergent lysate from equivalent numbers of siRNA duplexes transfected ST cells were subjected to reducing SDS-PAGE and immunoblotting with antibodies to p300, LC3, ATG5, ATG7 or β-actin. (b) ST cells were transfected with either specific siRNA duplexes 2# or control siRNA at the concentration of 100 nM for 24 h followed by infection with TGEV. At 24 hpi, cells were fixed and examined the TGEV infection by IFA. The percentage of TGEV-infected cells per view from three independent experiments is expressed as mean ± SD. (c) At 24 hpi, the extracellular virus titers in p300 siRNA 2#-transfected cells were measured by TCID₅₀. Representative data from three independent experiments are shown as the mean ± SD. *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. S9.