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. 2020 Mar 20;11:1254. doi: 10.1038/s41467-020-14992-9

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© The Author(s) 2020

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Fig. 4 — a Y2H analyses of interactions between BCM1 and CBEs. The transformed yeast strains were analyzed on selective medium lacking Leu and Trp (SD/-L-T) or His, Leu, Trp and Ura (SD/-H-L-T-U) in the presence of 30 mM 3-amino-1,2,4-triazole (3-AT). The combination of NubG-GluTR and Cub-GBP was used as the positive control. The NubG was used as the negative control for NubG-BCM1. b BiFC assays for interactions between BCM1 and CBEs. Co-expression of BCM1^ΔC20-YFPn with CHLH-YFPc served as the negative control, and the combination of GUN4-YFPn with CHLH-YFPc served as the positive control. Scale bars, 10 μm. c In vitro MgCh assay. Production of MgP in the assay was measured by HPLC and quantified relative to incubation time. Recombinant proteins used in the assay were stained with Coomassie Brilliant Blue (middle panel) and probed with GUN4 antibody (bottom panel), respectively. ND, not detected. Error bars represent SD of three biological replicates. Letters above histograms indicate significant differences as determined by Tukey’s HSD method (P < 0.05).