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tnf-α, il-1β, il–6 and il-8 gene expression in χ16022(pG8R8029) vaccinated and control zebrafish. Zebrafish were vaccinated with χ16022(pG8R8029) by i.c. injection. At three, five and seven days of post-immunization, total RNA was extracted from the gills, kidney, intestine and spleen and cDNA was prepared. The qRT-PCR assay was conducted to analyze the expression of tnf-α, il-1β, il-6 and il-8 genes using β-actin as an internal control. The results are expressed as mean ± standard error (bars) from three separate experiments. Differences between uninfected (control) and infected groups were analyzed by two-way ANOVA, where asterisks (*) indicate significant difference (*P < 0.05, **P < 0.01, ****P < 0.0001) with respect to the control group.
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