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. 2021 Aug 17;118(34):e2011900118. doi: 10.1073/pnas.2011900118

Fig. 1.

Fig. 1.

SINAT E3 ligases are positive regulators for BR-induced ethylene biosynthesis. (A) Y2H assays for the interaction between different ACS isoforms and SINAT E3 ligase isoforms. (B) BR-induced ethylene biosynthesis in dark-grown seedlings. Seedlings were grown on a medium with different concentrations of BR, and the accumulated ethylene was measured using gas chromatography. (C) BR-induced ethylene biosynthesis in seedlings expressing different levels of SINAT2 protein. (Inset) Expression levels of SINAT2 protein in independent transgenic lines expressing Myc-SINAT2 without BR treatment in the dark. The Myc-SINAT2 band intensity was normalized to HSC70 band, and the values were expressed relative to the ratio of Myc-SINAT2/HSC70 in no. 8, which was set to 1. (D) Co-IP of ACS5 and SINAT2 complex in N. benthamiana. The indicated combinations of tagged proteins were transiently expressed in N. benthamiana with or without BR treatment and used for co-IP analysis. GFP-ACS5 was immunoprecipitated with an anti-GFP antibody, and the resulting precipitates were further analyzed via immunoblotting using an anti-GFP or an anti-Myc antibody. Data represent the mean ± SD of three biological replicates (n = 3). *P < 0.05 and **P < 0.01 indicate a significant difference when samples were compared to the WT plants in each BR concentration using a Student’s t test.