Fig. 2: Evaluation of the interaction between GmCRY1b and DELLA proteins.
From: The mechanism of low blue light-induced leaf senescence mediated by GmCRY1s in soybean
a Schemes display full-length and truncated versions of the GmCRY1b protein. b Yeast two-hybrid (Y2H) assays show the interaction of the different truncated versions of GmCRY1b with DELLA proteins. SD-WL, minimal medium lacking Trp and Leu; SD-WLHA, selective medium lacking Trp, Leu, His, and adenine, and supplemented with 1 mM 3-AT. c Quantitation of β-galactosidase activity (Miller units) for each pair of bait and prey proteins as indicated by liquid assays. Values are mean ± SD (n = 3 biological replicates). The lowercase letters indicate significant differences (One-way ANOVA with two-sided Tukey test at a significance level of 0.05). d Co-IP assays demonstrate the blue light-dependent interaction of GmCRY1b with GmRGAa and GmRGAb in soybean leaves. Dark-adapted soybean leaves were either kept in the dark or exposed to blue light (50 μmol m−2 s−1) for 3 h. e BiFC analysis of protein interactions between GmCRY1b and GmRGAa in soybean protoplasts under WL or LBL conditions. The YFP fluorescence is shown, with the Merge representing the combination with the fluorescence of chloroplasts. Scale bars, 10 µM. The experiment was repeated three times with consistent results. f Effects of LBL treatment on the abundance of GmRGAb. Seedlings were grown under continuous WL for 12 days, then transferred into LBL or kept in continuous WL. The first trifoliate leaves were collected at the indicated time points. g The relative proteins levels of Flag-GmRGAb, normalized to HSP82 as in (f), are shown. Values are mean ± SD (n = 3 biological replicates). Statistical significance was analyzed using a two-tailed Student’s t-test, **P < 0.01, and P = 2.03 × 10−5, 5.57 × 10−8 and 2.82 × 10−8 at the time points of 3, 5 and 8 h. h Immunoblot analysis of transgenic GmRGAa and GmRGAb proteins in the indicated callus lines with anti-Flag antibody. i, j Relative transcript and protein levels of transgenic GmRGAa (i) and GmRGAb (j) in the indicated callus lines. Values are means ± SD (n = 3 biological replicates). Source data are provided as a Source Data file.
