Fig. 3 BIC1 and BIC2 inhibit early photoreactions of cryptochromes.

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Fig. 3 BIC1 and BIC2 inhibit early photoreactions of cryptochromes. BIC1 and BIC2 inhibit early photoreactions of cryptochromes. (A and B) BIC1 inhibits blue light–dependent phosphorylation of CRY1 and CRY2 and blue light–dependent degradation of CRY2. Immunoblots of samples prepared from 7-day-old etiolated seedlings (WT, wild type; +BIC1, BIC1-overexpressing) exposed to blue light (30 μmol m−2 s−1) for the indicated times were probed with antibodies to CRY1, CRY2, or HSP90 (loading control). Arrowheads indicate upshift bands of the phosphorylated CRYs. (C to E) Quantification of band intensities, showing CRY phosphorylation [(C), CRY1Pi/CRY1; (D), CRY2Pi/CRY2] or CRY2 degradation [(E), CRY2B/CRY2D]. (F) BIC inhibition of CRY2 photobodies. Protoplasts isolated from 4-week-old plants of indicated genotypes were transfected to express CRY2-YFP, exposed to blue light (20 μmol m−2s−1) for 0 to 60 min, fixed in 1% formaldehyde, and examined by a fluorescence microscope. Scale bar, 5 μm. (G) BiFC assay showing the blue light–independent (dispersed fluorescence) and blue light–dependent (photobodies) interactions between nYFP-CRY2 and cCFP-CRY2. Protoplasts transfected to express nYFP-CRY2 and cCFP-CRY2 were exposed to blue light and analyzed as in (F). (H) The CRY2-YFP photobody of the experiment shown in (F) was digitized and quantified; 30 nuclei per sample were counted, and the percentage of protoplasts containing photobodies was calculated (±SD; n = 3). (I) Same as (H) but for the nYFP-CRY2/cCFP-CRY2 photobody shown in (G). Qin Wang et al. Science 2016;354:343-347 Copyright © 2016, American Association for the Advancement of Science