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. 2021 Feb 4;22(4):1562. doi: 10.3390/ijms22041562

Figure 6.

Figure 6

OsSAUR33 interacts with OsSnRK1A and alters the expression of OsSnRK1A, OsMYBS1, and OsAmy3 during seed germination in rice. (A) Firefly luciferase (LUC) complementation imaging assay. nLUC-OsSnRK1A and cLUC-OsSAUR33 with the control vector were co-infiltrated into N. benthamiana leaves. LUC images were captured using a cooled Charge-coupled Device (CCD) imaging apparatus. (B) Bimolecular fluorescence complementation (BiFC) assay. OsSAUR33 and OsSnRK1A were fused to p2YC and p2YN, respectively. Different pairs of constructs were co-expressed in N. benthamiana. Yellow fluorescent protein (YFP) fluorescence was detected by confocal microscopy. Bar = 50 μm. (C) Maltose Binding Protein (MPB) pull-down assay of the OsSAUR33 and OsSnRK1A interaction. Anti-GST antibody was used to detect the output protein. The expression of OsSnRK1A (D,G), OsMYBS1 (E,H), and OsAmy3 (F,I) in the WT and the ossaur33 mutants at 12 h and 72 h of imbibition. The expression of genes was normalized to that of OsActin. Each column represents the mean ± standard deviation, black small symbol means the value of each replication, n = 3.