Extended Data Fig. 9: Investigation of the molecular mass of ScFT protein and evidence for its phosphorylation.
(a) Immunoblotting detection of ScFT in Weining rye leaf tissues at 7 days after sowing. Total leaf proteins (10 μg) were separated using 12% SDS-PAGE, followed by immunoblotting with an antibody against the peptide QLGRQTVYAPGWRQ conserved in ScFT1 and ScFT2. Based on the protein size markers, the ScFT protein detected (asterisk) had a molecular mass of ~29 kDa. (b) Detection of ScFT phosphorylation using Phos-tag SDS-PAGE coupled with immunoblotting. Total leaf proteins (20 μg), with or without a treatment by λ protein phosphatase for 30 min at 30 °C, were separated using 12% Phos-tag SDS-PAGE, followed by immunoblotting with ScFT specific antibody. A ScFT phosphoprotein band (arrow) was specifically detected for the sample that was not treated by λ protein phosphatase. The data shown were reproducible in three independent experiments.
