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. 2022 Apr 19;13:2055. doi: 10.1038/s41467-022-29648-z

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© The Author(s) 2022

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. 3 — a Transcriptional activation assay in yeast. Yeast cells were spotted on the indicated plates and grown at low (22 °C) or high (30 °C) temperatures for 96 h. BD, binding domain; AD, activation domain; SD-2, plates lacking Trp and Leu; SD-3 + 2 mM 3-AT, plates lacking Trp, Leu, and His, plus 2 mM 3-aminotriazole. b, c Protein levels of GFP-OsMS1 and GFP-OsMS1^wenmin1 are temperature-dependent in yeast cells (b) and N. benthamiana (c). N. benthamiana were incubated at 22 °C for 44 h and then transferred to the indicated temperatures for another 4 h before detecting the GFP signals. Red arrows indicate nuclei, white arrows indicate cytoplasm. d Quantitative analysis of (c). The GFP-OsMS1 and GFP-OsMS1^wenmin1 protein levels were quantitated and normalized to the mCherry level. P values indicate the significant differences relative to GFP-OsMS1 at 22 °C. e Dual-luciferase system. LUC, firefly luciferase. Ren-LUC, Renilla luciferase. f Relative luciferase activity of OsMS1-LUC and OsMS1^wenmin1-LUC in N. benthamiana plants during the low temperature to high-temperature transition. g OsMS1 and OsMS1^wenmin1 mRNA levels as in (f). h–j Protein levels of OsMS1-GUS and OsMS1^wenmin1-GUS are temperature-dependent in rice. pOsMS1:OsMS1-GUS and pOsMS1:OsMS1^wenmin1-GUS transgenic plants were grown at 22 °C or 30 °C in growth chamber. Protein levels of OsMS1-GUS and OsMS1^wenmin1-GUS were detected by GUS-staining of transgenic plant spikelets at stage 9 (h). OsMS1-GUS (i, left) and OsMS1^wenmin1-GUS (j, left) proteins were detected by ani-GUS antibody. Quantifications of OsMS1-GUS (i, right) and OsMS1^wenmin1-GUS (j, right) protein levels were relative to ACTIN. Scale bars, 5 μm (b), 50 μm (c), 1 mm (h). In d, f, g, i (right) and j (right). Data are presented as means ± s.e.m. (n = 6 biological replicates for d, n = 3 biological replicates for f, g, i, and j). Two-tailed unpaired t-test was used for statistical analysis. n.s., not significant. Source data are provided as a Source data file.