Extended Data Fig. 6: Oligomerization state of WT and mutated CRY2 in darkness and under blue light.
From: Structural insights into the photoactivation of Arabidopsis CRY2
a, Oligomerization state of wild type (WT) and mutated CRY2 measured by pull-down assays. Mammalian cells (Expi293F) were co-transfected with constructs designed to express the indicated proteins. The supernatant from lysed cells was incubated with StrepII beads under blue light (50 μmol m−2s−1) or in darkness for 1.5 h. Proteins in input and eluted fractions were detected by immunoblots probed with antibodies against StrepII and Myc. Residues critical for oligomerization are labeled in red. b, Quantification of results in a. The integrated densities of different bands were measured using Image J software. The pull-down prey bands under blue light were normalized by both input and bait bound. Mutants with pull-down bands lower than 20% percent of wild type were shown in red column. The standard deviation is derived from three measurements. Data were presented as mean ± SD. Black circles indicate individual data points for n=3 biological replicates. Differences between each mutants and wild type CRY2 were analyzed by two-tailed paired t-tests. Means with p < 0.05 are indicated. *, p < 0.05; **, p < 0.01; ***, p < 0.001; ****, p < 0.0001. Experiments in a were repeated three times, with similar results. Uncropped blot images are available as source data.
