Table 1.
Danio rerio primers and probes for quantitative PCR.
Figure 1.
Quantitative PCR of per1b, cry1b, per2 and cry1a in a Danio rerio embryonic cell line ZEM-2S.
The cells (2×10∧6) were stimulated with blue light (450–475 nm, 87.85 to 95.17 µwatts/cm2) for 10 min, and total RNA was extracted 1, 2, 6 and 12 h after the stimulus. ‘a’ is significantly different from ‘b’ and ‘b’ is significantly different from ‘c’ (p<0.05). In this and in figures 2 to 9, values are the mean ± Standard Error of Mean (n = 4–9).
Figure 2.
Quantitative PCR of per2 and cry1a in a Danio rerio embryonic cell line ZEM-2S.
The cells (2×10∧6) were stimulated with blue light (450–475 nm, 87.85 to 95.17 µwatts/cm2) for 10 min, in the presence or absence of the PLC inhibitor, U-73122, at 100 nM, and total RNA was extracted 2 h after the stimulus. In this and the following figures, the asterisk means statistically significant differences from all other groups (p<0.05).
Figure 3.
Quantitative PCR of per2 and cry1a in a Danio rerio embryonic cell line ZEM-2S.
The cells (2×10∧6) were stimulated with blue light (450–475 nm, 87.85 to 95.17 µwatts/cm2) for 10 min, in the presence or absence of the calcium chelator, BAPTA-AM, at 1 µM, and total RNA was extracted 2 h after the stimulus.
Figure 4.
Quantitative PCR of per2 and cry1a in a Danio rerio embryonic cell line ZEM-2S.
The cells (2×10∧6) were stimulated with blue light (450–475 nm, 87.85 to 95.17 µwatts/cm2) for 10 min, in the presence or absence of the PKC inhibitor, Ro 31-8220, at 100 nM and total RNA was extracted 2 h after the stimulus.
Figure 5.
Quantitative PCR of per2 and cry1a in a Danio rerio embryonic cell line ZEM-2S.
The cells (2×10∧6) were stimulated with blue light (450–475 nm, 87.85 to 95.17 µwatts/cm2) for 10 min, in the presence or absence of the CAMK II inhibitor, KN-93, at 1 µM, and total RNA was extracted 2 h after the stimulus.
Figure 6.
Quantitative PCR of per2 and cry1a in a Danio rerio embryonic cell line ZEM-2S.
The cells (2×10∧6) were stimulated with blue light (450–475 nm, 87.85 to 95.17 µwatts/cm2) for 10 min, in the presence or absence of the NOS inhibitor, L-NAME, at 1 mM and total RNA was extracted 2 h after the stimulus.
Figure 7.
Quantitative PCR of per2 and cry1a in a Danio rerio embryonic cell line ZEM-2S.
The cells (2×10∧6) were stimulated with the guanylyl cyclase activator, Y-C1, at 40 µM in DD, and total RNA was extracted after 2 h.
Figure 8.
Quantitative PCR of per2 and cry1a in a Danio rerio embryonic cell line ZEM-2S.
The cells (2×10∧6) were stimulated with blue light (450–475 nm, 87.85 to 95.17 µwatts/cm2) for 10 min, in the presence or absence of the MEK inhibitor, PD-98059 at 40 µM, and total RNA was extracted 2 h after the stimulus.
Figure 9.
Quantitative PCR of per2 and cry1a in a Danio rerio embryonic cell line ZEM-2S.
The cells (2×10∧6) were stimulated with blue light (450–475 nm, 87.85 to 95.17 µwatts/cm2) for 10 min, in the presence or absence of the adenylyl cyclase inhibitor, SQ-22536, at 20 µM, and total RNA was extracted 2 h after the stimulus.
Figure 10.
Quantification of cAMP in a Danio rerio embryonic cell line ZEM-2S.
The cells (8×10∧4) were stimulated with blue light (450–475 nm, 87.85 to 95.17 µwatts/cm2) for 1, 5 or 10 min, and cAMP was measured immediately after each light pulse. Forskolin at 10 µM was used as a positive control in DD. Values are the mean ± SEM (n = 3).