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Figure 6 | Veterinary Research

Figure 6

From: Antiviral activity of ISG15 against classical swine fever virus replication in porcine alveolar macrophages via inhibition of autophagy by ISGylating BECN1

Figure 6

Autophagy protein BECN1 is ISGylated by ISG15.A–C Influence of ISG15 on autophagy and CSFV replication in BECN1-overexpressing cells. Expression of LC3 protein (A) and the number of LC3 puncta per cell (B) were determined by Western blot and image analysis, respectively. C CSFV replication was quantified by RT-qPCR and IFA, respectively. D ISGylation of BECN1 was detected in cells transfected with 3.1, 3.1-ISG15AA, and 3.1-ISG15GG plasmids. PAMs were transfected with 3.1, Flag-ISG15, Flag-BECN1, and Flag-ISG15-BECN1 and then infected with CSFV. E Expression of LC3 protein and fusion protein was probed by Western blot with an anti-LC3 antibody and an anti-Flag antibody, respectively. F The number of LC3 puncta per cell was determined by image analysis. G CSFV replication in these cells was quantified by RT-qPCR and IFA, respectively. Data (B, C, F, and G) represent the mean ± SD of three independent experiments and were measured in technical duplicate. Comparisons between groups were calculated using Student’s t-test. *P < 0.05; **P < 0.01; ***P < 0.001

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Veterinary Research

ISSN: 1297-9716