FIG. 6.

Quantitative gene expression of Sox9a and Col2A1 in medaka caudal peduncle. Medaka embryos were exposed to DMSO or a graded concentration of TCDD (0.05–1 ppb) at 4 hpf for 1 h, washed, and incubated in ERM at 26°C. At 10 dpf, embryos were dechorionated and caudal peduncles processed for RNA and gene expression as described in “Materials and Methods.” (A) Relative Sox9a gene expression, (B) relative Col2A1 1 of 2 gene expression, and (C) relative Col2A1 2 of 2 gene expression. mRNA transcript levels were normalized to medaka 18s and are represented as the mean mRNA level ± SEM. A significant change in transcript was determined using one-way ANOVA, p value < 0.05, followed by Newman-Keuls Multiple Comparison test within the Prism4 software package (GraphPad Software Inc., San Diego, CA). Data were logarithmically transformed as needed to improve equality of variances. All experiments consisted of three replicate experimental wells containing 10 embryo caudal peduncles per well and each experiment was replicated at least three times. All qPCR was conducted in triplicate. Different letters indicate significant differences (p < 0.05) between fields.