Analysis of p38 MAPK activation and inflammatory cytokines expression during infection with recombinant SARS-CoV-∆E virus after passage in mice.

<p>(A) Vero E6 cells were mock-infected or infected with the wt, ∆E and ∆E-8a-dup viruses at a moi of 0.3 and the presence of active phosphorylated (p-p38) and total (p38) p38 MAPK was detected by Western blot analysis at 24 hpi. Actin was used as control. Lines 1, 2 and 3 indicated three independent experiments analyzed in each case. (B) Phospho and total p38 MAPK amounts were quantified by densitometric analysis. The graph shows the phosphorylated p38/total p38 MAPK ratio in Vero E6 cells infected with wt, ΔE and ∆E-8a-dup viruses at 24 hpi. Statistically significant data compared to mock-infected cells are indicated with one (P < 0.05) or two (P < 0.01) asterisks. (C) Expression of inflammatory cytokines in lungs of infected mice evaluated by RT-qPCR at 2 dpi. Three independent experiments were analyzed with similar results in all cases. Error bars represent the means of three experiments analyzed for each condition. (D) Vero E6 cells were mock-infected or infected with the wt or ∆E-8a-dup viruses at a moi of 0.3 and the presence of active phosphorylated (p-p38) and total (p38) p38 MAPK was detected by Western blot analysis at 24 hpi. Actin was used as control. Lines 1, 2 and 3 indicated three independent experiments analyzed in each case. (E) Phospho and total p38 MAPK amounts were quantified by densitometric analysis. The graph shows the phosphorylated p38/total p38 MAPK ratio in Vero E6 cells infected with wt and ∆E-8a-dup viruses at 24 hpi. Statistically significant data compared to mock-infected cells are indicated with one (P < 0.05) asterisk. (F) Expression of inflammatory cytokines in lungs of infected mice evaluated by RT-qPCR at 2 dpi. Three independent experiments were analyzed with similar results in all cases. Error bars represent the means of three experiments analyzed for each condition.