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. 2021 May 2;33(7):2320–2339. doi: 10.1093/plcell/koab120

Figure 6.

Figure 6

SlPIF4 is required for SlDYT1-mediated transcriptional activation of SlTDF1 under moderately low temperatures. A, Interaction of SlPIF4 with SlDYT1 in Y2H assays. Protein–protein interactions were assessed on SD/-Ade/-His/-Trp/-Leu (SD/-4) medium containing 10 mM 3-amino-1,2,4-triazole (AT). Empty pGBKT7-53 (BD-53) and pGADT7-T1 (AD) vectors were included as the positive control. Empty pGBKT7-Lam and pGADT7-T1 vectors were included as the negative control. B, BiFC confocal images showing SlPIF4 and SlDYT1 interaction in vivo. Nuclear YFP fluorescence was detected in N. benthamiana leaves expressing SlPIF4 fused to the YFP N-terminal region (2YN) and SlDYT1 fused to the YFP C-terminal region (2YC). Empty 2YN and 2YC vectors were included as the positive control. The positions of nuclei are shown by the H2B-RFP marker. Bars = 50 µm. C, SlPIF4 enhances SlDYT1-mediated transcriptional activation of SlTDF1 under moderately low temperature conditions. The LUC/REN mean value of the empty vector pSlTDF1-LUC was set to 1 as the calibrator. Vector, empty vector pSlTDF1-LUC; NT, normal temperature (25°C/20°C, day/night for 3 d); MLT (25°C/20°C, day/night for 2 d, followed by 10°C for 24 h). Each error bar represents the mean ± se, n = 6–8 biological replicates. Asterisks indicate significant differences: **P < 0.05 (Student’s t test).