Fig. 4: ZmCRY1c interacts with GLOSSY2 in a blue light-strengthened manner.

a Histidine auxotrophy assays showing the interaction between ZmCRY1c and GL2 in a blue light-strengthened manner. Yeast cells were grown at –LWHA medium in the dark or under blue light (20 μmol m^–2 s^–1). P53/T was a positive control pair. b β-gal assays of yeast cells grown on –LW medium at 28 °C in darkness (D) or under blue light (B, 20 μmol m^–2 s^–1) for the indicated times. Error bars represent the SD of three independent biological replicates. Data are presented as mean values ± SD. ZmCRY1c^D381A is a site-specific mutant of ZmCRY1c that cannot be activated by blue light. c Co-localization of ZmCRY1c and GL2 in the cytoplasm in maize protoplasts. BF, brightfield. Merge, overlay of the GFP, CFP, and brightfield images. Scale bar = 10 μm. d Gray-value analysis of fluorescent signals along the dashed diagonal line in the left images. e, f BiLC and BIFC assays showing in vivo protein interactions between ZmCRY1c and GL2. Leaf epidermal cells of Nicotiana benthamiana were co-transformed to express the indicated fusion proteins. In BIFC assay, ZmCRY1c^D381A-cCFP and nYFP-KCS4 served as negative controls. Scale bar = 200 μm. g Co-IP assays showing that ZmCRY1c interacts with GL2 in a blue light-strengthened manner in plant cells. Seven-day-old 28 °C LD-grown seedlings were pretreated in darkness for 48 h, then exposed to blue light (B, 20 μmol m^–2 s^–1) for 2 h. Input: immunoblots showing the abundance of GL2 and YFP-ZmCRY1c in total protein extracts. GL2 immunoprecipitation (IP): IP products precipitated by the anti-GL2 antibody. Total proteins (Input) or IP products of GL2-beads (GL2 IP) were probed in immunoblots with an anti-GL2 or anti-GFP antibody. In a and c–g, three independent experiments were performed with similar results.