Figure 2 | Scientific Reports

Figure 2

From: Gpr19 is a circadian clock-controlled orphan GPCR with a role in modulating free-running period and light resetting capacity of the circadian clock

Figure 2

The Gpr19 CRE sequence has the ability to produce circadian transcription in the SCN. (A) The CRE in the Gpr19 promoter. Genomic positions relative to the transcription start site (+ 1) of the Gpr19 gene are indicated along with evolutionary conservation scores among mammalian species. Alignment shows the CRE (− 867 to − 860; highlighted in magenta) and its flanking sequences of mouse, human, and other representative mammalian species. We used reporter constructs containing serial deletions of the mouse Gpr19 promoter (− 242 to + 226, − 514 to + 226, − 915 to + 226, − 1083 to + 226) and the mutant derivative for the CRE (mut; GCACAAAA). We also used reporter constructs containing 3 × isolated CRE (3 × CREwt) or its mutant (3 × CREmut). miniP, minimal promoter. (B) Gpr19 promoter activities in MEF cells after treatment with cAMP enhancer FSK. Average fold increase relative to basal activity was calculated (n = 6, for each construct). Error bars indicate SEM. ***P < 0.001, one-way ANOVA, Bonferroni’s post hoc test. (C) Representative detrended bioluminescence traces from SCN explants infected with AAV carrying the 3 × CREwt (upper) or 3 × CREmut (lower) reporter construct. Luminescence was recorded at 20-min intervals over 5 days in culture.

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