Figure 8
Expression of clock genes and the D-box reporter construct in cavefish and zebrafish cells. qRT-PCR analysis of (A,C) cry1a and (B,D) per2 in cells exposed for 9 hours to (A,B) blue light and (C,D) 300 μM H2O2. Mean mRNA relative expression (n = 3) ± SD is plotted on the y-axis, whereas time is plotted on the x-axis. Statistical tests of the differences in kinetics of gene expression between the two cell lines were analyzed with two-way ANOVA followed by Bonferroni multiple comparable T-test. Statistically significant differences between peaks of expression are indicated (*p < 0.05; **p < 0,01; ***p < 0,001). (E) Representative real time luciferase assay of PAC-2 (red trace) and EPA cells (black trace) transfected with the D-boxcry1a-Luc reporter and treated with H2O2 and blue light (left and right sides of the panel, respectively). Means of bioluminescence (CPS) (n = 8 wells) are plotted on the y-axis and the extent of the dark and blue light periods are indicated on the x-axis. See Fig. S2B for controls. (F) DCF-DA assay of EPA cells during 4 hours of exposure to blue (blue bars) or red (red bars) light. The mean of fold induction ± SD with respect to time 0 (n = 24) are plotted on the y-axis and time on the x axis. Levels of significance between peak points of expression are indicated (***p < 0.001, **p < 0.01, *p < 0.05).
