Syncrip facilitates porcine parvovirus viral dna replication through the alternative splicing of ns1 mrna to promote ns2 mrna formation
Figure 3 PPV infection up‑regulates SYNCRIP expression in vitro and in vivo
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| Figure 3 PPV infection up‑regulates SYNCRIP expression in vitro and in vivo. PK-15 cells were infected with 1 MOI of PPV or mock infection
for the time indicated, the mRNA level of SYNCRIP (A), the protein level of SYNCRIP (B) and the relative viral titers of different infection times were measured by TCID 50 (C). D‑E qPCR and western blotting analysis of SYNCRIP mRNA and protein levels in PK-15 cells infected with different doses of PPV (MOI = 0, 0.5, 1, or 2) for 24 h. β-actin served as an internal control. F–G Western blotting and qPCR analysis of SYNCRIP protein and mRNA levels in different tissues of PPV infected pregnant sows. H–I Western blotting and q-PCR analysis of viral VP2 protein levels and DNA copies in different tissues of PPV infected pregnant sows. β-actin served as an internal control. The results are shown as the mean ± SD (n = 3). *p < 0.05, versus mock infected cells at same time points. # p < 0.05, versus mock tissues at same tissue. Page 8 of 15 Chen et al. Vet Res (2021) 52:73 4) for 24 h. The results show that SYNCRIP expression significantly increased with the doses of PPV infection, and the expression level of SYNCRIP is associated with viral infection level (Figures 3 D, E). These results dem- onstrate that the expression of SYNCRIP is upregulated by PPV infection in vitro. To further confirm whether the expression of SYNCRIP is up-regulated by PPV infection in vivo, we examined the mRNA and protein levels of SYNCRIP in different tissues of pregnant sows infected by PPV. The results show that the mRNA level of SYNCRIP in major targeted tissues (spleen, lung, kid- ney, ovary and uterus) increased compared to those in the nonpathological tissues (heart, liver and brain) of PPV-infected pregnant sows at 35 days post-infection (dpi) (Figure 3 F). Consistently, SYNCRIP protein lev- els varied in different tissues of control pregnant sows and also apparently increased in lung, kidney, ovary and uterus of PPV-infected pregnant sows at 35 days post- infection (Figure 3 G). Simultaneously, PPV VP2 protein levels and viral loads were higher in major pathological tissues (spleen, lung, kidney, ovary and uterus) than those in the nonpathological tissues (heart, liver and brain) of PPV-infected pregnant sows (Figures 3 H, I). These results demonstrate that the expression of SYNCRIP was regu- lated in PPV-infected cells and tissues. tải về 5.65 Mb. Chia sẻ với bạn bè của bạn:
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