CRY1 expression is not temperature regulated. (A) qPCR results showing the lack of dramatic change of CRY1 transcription in response to increased temperature. Four-day-old seedlings grown in 22 °C in continuous white light condition were transferred to 28 °C or were continually placed at 22 °C for a 5-h time course, respectively. Error bars represent SD of three technical replicates of qPCR. Each experiment was performed at least three times with similar results. (B and C) Immunoblots showing the lack of temperature effects on the abundance of the CRY1 proteins in 6-d-old seedlings under the conditions tested. Samples were fractionated by 10% (wt/vol) Acrylamide SDS/PAGE, blotted, and probed with the anti-CRY1 antibody. WT plants were grown in 22 °C long-day (LD) condition for 6 d, transferred to 16 °C continuous light condition (B), or to 28 °C (C) for 16 h, then transferred to 28 °C (B) or 16 °C (C) for the indicated time before sample collection.