Characterization of GmSIN1. Characterization of GmSIN1. (A) GmSIN1-GFP fusion proteins localized to the nucleus in transiently transformed Arabidopsis protoplasts. (Left) Images of a protoplast harboring p35Spro:GmSIN1-GFP and pMDC32-1A BES1n-mCherry (nuclear marker). (Right) Images of a protoplast harboring p35Spro:GFP. The top row shows the GFP signal (green), the middle row shows the nuclear marker (left) and chloroplast autofluorescence (right; magenta), and the bottom row shows the merged images. The images show representative results from more than 30 transformed protoplasts from three independent experiments. More images can be found in Supplemental Figure 8B. Bar = 20 μm. (B) Ability of yeast transformants to grow on medium lacking His and Leu but containing 10 mM 3-aminotriazole, and the formation of color in the X-β-Gal assay indicates transcriptional activation. The images show representative results from more than five independent yeast transformants. BD, Yeast colony expressing GAL4 DNA binding domain; BD-GmSIN1, Yeast colony expressing BD-GmSIN1 fusion protein. (C) Expression of GmSIN1 analyzed using RT-qPCR in root, stem, leaf, flower, and seed tissue in cv Shengdou No. 9. (D) Localization of GmSIN1 mRNA using in situ hybridization. (Left) Transverse section of a root 1 mm from the root tip of 6-d-old cv Shengdou No. 9 plants probed with a GmSIN1 antisense probe. (Right) Profile obtained using a sense strand probe. Bar = 100 μm. (E) Expression of GmSIN1 analyzed using RT-qPCR in response to NaCl (150 mM), moisture stress (20% [w/v] polyethylene glycol [PEG] 6000), low temperature (4°C), 100 µM ABA, or 1 mM H2O2 treatment in cv Shengdou No.9. The roots of 2-week-old seedlings were collected for RNA extraction. Error bars represent se (n = 3 biological repeats). The details of the sampling methods are shown in “Methods.” Shuo Li et al. Plant Cell 2019;31:2107-2130 ©2019 by American Society of Plant Biologists