The Arabidopsis protein extraction was performed according to Saucedo et al. (2019) (link) with modifications. Briefly, 400 mg of fresh tissue of entire 2-week-old Arabidopsis plants were pulverized with liquid nitrogen and then 4 mL of extraction buffer [0.5 M Tris–HCl pH 8, 0.7 M Sucrose, 2 protease inhibitor tablets (complete Mini, EDTA-free-Thermo Scientific Lot # UG27666820), 50- mM EDTA, KCl 0.1 M, β-mercaptoethanol 0.2%)] were added and homogenized until the sample was completely thawed. Then, 2 mL of basic saturated phenol pH 8.0 (Winkler, Hackensack, NJ, USA; # 20192088) were added, shaken 10 min in ice, and centrifuged at 8,000 g for 19 min at 4°C. The supernatant was recovered and four volumes of 0.1-M ammonium acetate in methanol (Merck gradient for liquid chromatography) were added, and proteins were precipitate overnight at −20°C. After centrifugation, the pellet was washed with 0.1-M ammonium acetate in methanol, and with acetone. The remaining acetone was evaporated at room temperature and the proteins (pellet) were stored at −20°C in resuspension buffer (100-mM Tris–HCl pH 7.0, 1% SDS). The extraction was carried out in triplicate for each transgenic line and control.