Expression profiles of pro-inflammatory and stress-inducible genes.

<p>The graphs present the mRNA expression of (A) interleukin 1β (<i>IL-1β</i>), (B) interleukin 6 (<i>IL-6</i>), (C) interleukin 8 (<i>IL-8</i>), (D) interleukin 10 (<i>IL-10</i>), (E) macrophage receptor MARCO (<i>MARCO</i>), (F) activating transcription factor 3 (<i>ATF3</i>), (G) tumor necrosis factor (<i>TNFα</i>), (H) C-X-C motif chemokine receptor 1 (<i>CXCR1</i>) and (I) C-X-C motif chemokine receptor 2 (<i>CXCR2</i>) from pooled (n = 30) 5dpf wild-type zebrafish larvae in four experimental groups: 1) control, 2) exposed to 10µM CuSO₄ for 40 min, 3) exposed to a mixture of 100 nM PACAP-38 + 10µM CuSO₄ for 40 min preceded by one hour pre-incubation with 100 nM PACAP-38 only, and 4) exposed to 100nM PACAP-38 only. Each group was covered by samples analyzed in triplicate in three separate experiments. Data in the figure represent the average of the three individual experiments. Gene expression values were normalized to housekeeping gene <i>β-actin</i>. To maintain image clarity, only differences between CuSO₄ exposed and PACAP-38 co-treated groups are marked on the graphs. Co-treatment with PACAP-38 significantly reduced up-regulated by copper treatment <i>IL-1β</i> and <i>IL-6</i>, <i>IL-8</i> and <i>ATF3</i> gene expressions (A, B, C and F) (one-way ANOVA, Kruskal–Wallis test with Dunn’s posttest, GraphPad Prism 5, <i>p</i> < 0.05), while it had no significant influence on <i>IL-10</i>, <i>MARCO</i> or <i>TNFα</i> genes (D, E and G) (one-way ANOVA, Kruskal–Wallis test with Dunn’s posttest, GraphPad Prism 5, <i>p</i> > 0.05). With regard to <i>CXCR1</i> and <i>CXCR2</i>, 10 µM copper chemical injury and 100 nM PACAP-38 treatment did not significantly change the expression level of the receptors. (one-way ANOVA, Kruskal–Wallis test with Dunn’s post-test, GraphPad Prism 5, <i>p</i> > 0.05).