Fig. 2.

JARID1a is required for normal circadian function. (A) Real-time bioluminescence from Jarid1a^+/+ (black) and Jarid1a^−/− (red) mouse fibroblasts stably expressing a Per2:Luc construct. (Middle and right) Average period lengths (+SEM, n = 10) and amplitude estimates (***P < 0.001, two-tailed Student’s t test). (B) Per2, Cry1, Dbp, and SDF1 mRNA abundance in Jarid1a^+/+cells (black) and Jarid1a^−/− cells (red). Forty-eight hours after circadian synchronization, cells were collected at 3-hour intervals over the course of 24 hours, and the mRNA abundance was analyzed by reverse transcription (RT)-QPCR and normalized to that of Gapdh. Average normalized values (±SEM, n = 3) are plotted at various times after cell synchronization. (C) Representative real-time luminescence from U2OS cells stably expressing Bmal1:Luc and transfected with scrambled (S, black) or Jarid1a siRNA (red). Effects of different Jarid1a siRNA concentrations (10, 20, and 40 nM) on the endogenous Jarid1a mRNA abundance (average + SEM, n = 3) and period length (average + SEM, n = 5) relative to those of cells transfected with scrambled siRNA (40 nM). Representative results from at least three independent repetitions are shown.