The transcription and translation of the transcriptional repressors of the circadian clock are synchronous. (A, top) The average CRY2 band intensity from three protein immunoblots in wild-type synchronized U2OS cells is plotted as a function of time. All measurements were normalized to beta-actin. Error bars represent standard error of the mean. mRNA abundance measurements for CRY2 from total RNA-seq are plotted alongside for comparison. This is the same line displayed in part C of this figure. (A, bottom) Representative protein immunoblots for CRY2 and beta-actin are shown. (B) The lag in time between peak RPF and peak mRNA level is plotted for all genes that oscillate in both RNA and RPF data. Phases were calculated using JTK_CYCLE. (C) RNA and RPF traces for the negative regulators of the circadian clock are plotted as a function of time. Blue traces for the RNA data use the axes on the left. Red traces for the RPF data use the axes on the right. Points from both replicates are displayed, and the lines are plotted using a moving average (see Supplemental Methods for further detail). Traces from siARNTL libraries are displayed in Supplemental Figure S9.
