SB203580 treatment reduces GGT expression in DENV-infected mice.

<p>Mice were infected with 4 × 10⁵ FFU/ml of DENV and treated with 2%DMSO (v/v) or SB203580 dissolved in 2%DMSO. The control (uninfected) group was treated with 2%DMSO (v/v) alone. Treatments were given 1 h before and after DENV infection and again at 24 h after infection. On day 7 after infection, the liver tissues were collected, and the proteins were extracted for western blotting analysis using antibody directed against (A) GGT normalized to GAPDH. The results shown are representatives of three independent experiments with three mice (n = 3) from each group. A densitometry analysis using the ImageJ software is shown in (B).</p

SB203580 treatment reduces caspase 3 expression.

<p>Mice were infected with 4 × 10⁵ FFU/ml of DENV and treated with 2%DMSO (v/v) or SB203580 dissolved in 2%DMSO. The control (un-infected) group was treated with 2%DMSO (v/v) alone. Treatments were given 1 h before and after DENV infection and again at 24 h after infection. On day 7 after infection, the liver tissues were collected, and the proteins were extracted for western blotting analysis using antibodies directed against (A) pro caspase 3 and (C) cleaved caspase 3. The results shown are representative of three independent experiments with three mice (n = 3) from each group. A densitometry analysis using the ImageJ software is shown in (B) pro caspase 3 and (D) cleaved caspase 3.</p

Apoptotic gene expression profiles of DMSO-treated DENV-infected mice and SB203580 treated DENV-infected mice (both are normalized with those of DMSO-treated uninfected mice).

<p>Apoptotic gene expression profiles of DMSO-treated DENV-infected mice and SB203580 treated DENV-infected mice (both are normalized with those of DMSO-treated uninfected mice).</p

SB203580 treatment reduces caspase 8 expression.

<p>Mice were infected with 4 × 10⁵ FFU/ml of DENV and treated with 2%DMSO (v/v) or SB203580 dissolved in 2%DMSO. The control (uninfected) group was treated with 2%DMSO (v/v) alone. Treatments were given 1 h before and after DENV infection and again at 24 h after infection. On day 7 after infection, the liver tissues were collected, and the proteins were extracted for western blotting analysis using antibody directed against the pro and p18 subunit of the cleaved form of (A) caspase 8. The results shown are representative of three independent experiments with three mice (n = 3) from each group. A densitometry analysis using the ImageJ software is shown in (B).</p

SB203580 does not reduce the phosphorylation of p38 MAPK.

<p>Proteins were extracted from the liver tissue samples of 2%DMSO-treated (uninfected), 2%DMSO-treated DENV-infected, and SB203580-treated DENV-infected groups of mice. An additional cocktail of phosphatase inhibitors was added for maintaining the phosphorylated proteins and subjected to western blot analysis with specific antibodies. Results were shown for (A) phosphorylated p38 MAPK and (B) total p38 MAPK. The results shown are representative of three independent experiments with three mice (n = 3) from each group. A densitometry analysis was conducted for the individual blots, normalized to the respective GAPDH, is shown in (C) phosphorylated p38 MAPK and total p38MAPK.</p

SB203580 treatment reduces the phosphorylation of MAPKAPK2 and HSP27.

<p>Proteins were extracted from the liver tissue samples of 2%DMSO-treated (un-infected), 2%DMSO-treated DENV-infected, and SB203580-treated DENV-infected groups of mice. An additional cocktail of phosphatase inhibitors was added for maintaining the phosphorylated proteins and allowed them to standard Western blot analysis with specific antibodies. The results were shown (A) phosphorylated MAPKAPK2 and total MAPKAPK2 and (C) phosphorylated HSP27 and total HSP27, which is normalized to their respective GAPDH. The results shown are representative of three independent experiments with three mice (n = 3) from each group. A densitometry analysis was conducted with the individual blots normalized to GAPDH and is shown in (B) phosphorylated MAPKAPK2 and total MAPKAPK2 and (D) phosphorylated HSP27 and total HSP27.</p

The hematological phenotype of the DENV-infected mice was improved by SB203580 treatment.

<p>Mice were infected with DENV and treated with 2%DMSO or SB203580. The uninfected control group was also treated with 2%DMSO alone. Seven days after infection, their blood was collected in EDTA tubes for hematological analysis. (A) WBC counts, and (B) platelet counts. The results were pooled from two independent experiments, and are presented as means ± SEM (a total of 12 mice per group from two independent experiments). The asterisks indicate statistically significant differences between groups (<i>p</i> < 0.05).</p

SB203580 treatment reduces the phosphorylation of ATF-2.

<p>Proteins were extracted from the liver tissue samples of 2%DMSO-treated (uninfected), 2%DMSO-treated DENV-infected, and SB203580-treated DENV-infected groups of mice. An additional cocktail of phosphatase inhibitors was added for maintaining the phosphorylated proteins and allowed them to standard Western blot analysis with specific antibodies. The result is shown (A) phosphorylated ATF-2 and total ATF-2 normalized to GAPDH. The results shown are representative of three independent experiments with three mice (n = 3) from each group. A densitometry analysis is conducted using the ImageJ software and is shown in (B).</p

SB203580 treatment reduces liver enzymes in DENV-infected mice.

<p>Mice were infected with DENV and treated with 2%DMSO or SB203580. The uninfected control group was also treated with 2%DMSO. Seven days after infection, blood was collected and sera prepared to estimate liver transaminases. (A) ALT, and (B) AST. The pooled results of two independent experiments are presented as means ± SEM (12 mice per group in two independent experiments). The asterisks indicate statistically significant differences between the groups (<i>p</i> < 0.05).</p

SB203580 modulates the cytokine and chemokine gene expressions in DENV infection

<p>Mice were infected with 4 × 10⁵ FFU/ml of DENV and treated with 2%DMSO (v/v) or SB203580 dissolved in 2%DMSO. The control (uninfected) group was treated with 2%DMSO (v/v) alone. Treatments were given 1 h before and after DENV infection and again at 24 h after infection. On day 7 after infection, the liver tissues were collected, and RNA was extracted, cDNA were prepared and which undergone Real-time RT PCR analysis with individual primer set. GAPDH is used as the house keeping gene. The mRNA expression of (A) TNF-α (B) IL-6 (C) IL-10 (D) CCL-5 (E) CXCL-10 are shown. Results were represented in the graph by three independent experiments for at least 3 independent mice from each group. Statistical analysis is conducted by One Way ANOVA using GraphPad Prism Software Version 5. The asterisks indicate statistically significant differences between groups (<i>p</i>< 0.05).</p