SUR-8 in clock cells regulates PER abundance.
a Western blots of head extracts from SUR-8 downregulation at indicated time on 1st day of DD. Membranes were probed with anti-PER and anti-Lamin antibodies. Lamin was used as a loading control. * indicates the nonspecific band. C for control group, TD2/+; KD for experimental group, TD2/+;Sur-8RNAi-1/+. Quantification of PER levels is shown on the right. PER protein levels are normalized to Lamin. PER levels at CT1 in TD2/+ is set to1. The quantification curve represents three independent repeats. Error bars indicate SEM. *P < 0.05, **P < 0.01, unpaired t-test. b Western blots of head extracts from Sur-8 overexpression at indicated time on 1st day of DD. The band labeled with * is a nonspecific band. C for control group, UAS-Sur8-Flag; OE for experimental group, UAS-Sur8-Flag;TG4-16. Quantification of PER levels is shown on the right. Error bars indicate SEM. *P < 0.05, unpaired t-test. c Immunostaining of sLNvs for PER (red) and PDF (green) on the 5th day in DD. Fly brains were fixed, dissected at 4-hour intervals, and imaged under Leica SP8 confocal microscopy. Same parameters were applied to image all fly brains. At each timepoint, at least 15 brains from three independent experiments were quantified. Gray bar, subjective day; dark bar, subjective night.