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976364.fig.004b
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Figure 4: Mice were anesthetized and then infected intranasally by dropping 0.05 mL of influenza virus suspension (4LD50) except normal control group. High-dose SFXF (3.76 mg/kg), medium-dose SFXF (1.88 mg/kg), low-dose SFXF (0.94 mg/kg), or Oseltamivir (11.375 mg/kg) was administrated daily starting at 2 hours before the first viral infection until 4 days post-infection (4 dpi). Then total RNA was isolated from lung tissues of normal control, virus control, and Oseltamivir- and SFXF-treated mice and was analyzed by real-time PCR and western immunoblotting. (a) Relative quantification of the TLR7, MyD88, JNK, and p38 mRNA expressed in the mice lung tissue of six groups, with 12 mice in each group. The quantity of mouse TLR7, MyD88, JNK, and p38 mRNA expression was normalized to the mRNA expression of mouse GAPDH (a housekeeping gene). The quantities are shown as mean ± standard deviation (SD) and two indicated groups as determined by the Newman-Keuls multiple comparison test following one-way ANOVA. Compared with the normal control group: ## and compared with the virus control group: * , ** . (b) Western immunoblottings of TLR7, MyD88, JNK, and p38 protein expressions in the mice lung tissue of six groups, with 12 mice in each group. In normal control group, mice were blank and not infected with H1N1 virus (N). Mice were infected with H1N1 virus but not treated with any drugs (M). And mice were infected with H1N1 virus with treatment of high-dose SFXF (SH), medium-dose SFXF (SM), low-dose SFXF (SL), and Oseltamivir. The total gray value of each band was determined using ECL reagents and IPP software. (c) Western immunoblottings of relative quantification of TLR7, MyD88, JNK, and p38 protein levels. The relative intensity data were shown as the ratios of the target gene intensity to GAPDH intensity. Data shown were the mean ± SD of three independent experiments. * and ** versus the virus control group (virus only).