Figure 3
From: Melanopsin, a Canonical Light Receptor, Mediates Thermal Activation of Clock Genes
Expression of Per1 (A and C) and Bmal1 (B and D) in murine Melan-a melanocytes and B16-F10 melanoma cells after heat stimulus (39.5 °C) in the presence of AA92593. Melan-a or B16-F10 cells were kept for 3 days in constant dark and temperature (37 °C). In the beginning of the 4th day, Melan-a cells were synchronized by two medium changes, and after further 24 hours cells were heat-stimulated. For B16-F10 cells, the heat shock (39.5 °C) was applied at the beginning of the 4th day. In both scenarios, cells were divided into four groups: (1) control group at 37 °C in the presence of DMSO (0.1%); (2) heat-stimulated (39.5 °C) group in the presence of DMSO (0.1%); (3) group kept at 37 °C in the presence of AA92593 (10 µM), a selective OPN4 antagonist; (4) heat-stimulated (39.5 °C) group in the presence of AA92593 (10 µM). Total RNA was extracted immediately and 1 h after the end of the stimulus for Melan-a and B16-F10 cells, respectively. Boxplots show the median, quartiles, maximum, and minimum expression values of each gene transcript normalized by Rpl 37a and expressed relative to the minimal value of DMSO group at 37 °C (N = 5–9). Statistical analysis was performed by One-way ANOVA followed by Tukey’s test.
