Figure 4.

Expression of genes associated with eicosanoid synthesis and prostaglandin E₂ production in zebrafish kidneys. (A–D) Real-time PCR quantification of expression of selected genes involved in eicosanoid synthesis in zebrafish kidneys. Data are presented as ΔCT (delta cycle threshold, expression normalized to housekeeping gene EF1) using box and whisker plots. Expression of genes before and after challenge is represented by solid colored and pattern-filled boxes, respectively (n = 4 biological replicates, a pool of five fish each). Significant difference between treatments before and after challenge is indicated in asterisk bracket with corresponding p value, as determined by t-test. A and B denote significant differences between treatments before challenge; a and b denote significant differences between treatments after challenge. Differences were considered significant at p < 0.05. (E) Prostaglandin E₂ levels in kidneys of zebrafish. ELISA was used to quantify PGE₂ concentration in homogenized kidney samples obtained from zebrafish fed with different diets. Box plot represents average pg PGE₂/μg protein ± SD (n = 15, 3 biological replicates/treatment) for each dietary treatment. PGE₂ levels before and after challenge represented by solid-colored and pattern-filled boxes, respectively. Significant difference (p-values of t-test) between treatments before and after challenge is indicated by bracket.