Fig. 2.
Oxidation state of PRX in constant conditions in C. elegans. (A) Representative immunoblot probed for over-/hyper-oxidized 2-Cys PRX (PRX-SO2/3). After entrainment to a temperature cycle (16 h at 13 °C and 8 h at 16 °C, darkness), the oxidation of PRDX-2 shows a circadian rhythm in free-running conditions (13 °C, darkness). Time point 0 h corresponds to the release from warm to cold. Loading control shows an image of the same blot probed for β-actin. For each time series, the data were quantified by densitometry and normalized to the highest value within each blot. The x axes are labeled as described in Fig. 1B. The mean ± SEM for three biological replicates is shown. A sinusoidal curve was fitted to the data using Circwave (P < 0.01). Additional statistical analysis with Circwave and JTK-Cycle is shown in Table S1. (B) prdx-2 mRNA levels are not rhythmic under the same protocol used for protein quantification. The expression of prdx-2 was measured by quantitative RT-PCR. The results were normalized to the amount of act-4 mRNA and then to the sample with the highest expression level. The x axes are labeled as described in Fig. 1B. The plot shows the mean ± SEM of three biological replicates.