Fig. 1
From: Maternal control of suspensor programmed cell death via gibberellin signaling
NtCRF1 binds to the NtCYS promoter. a Distribution of AATTT motifs (M) in the promoter region (−1.4 kb) of NtCYS. Fragments I (−189 to −83 bp), II (−396 to −296 bp), III (−609 to −476 bp), and IV (−1300 to −1211 bp) were subjected to ChIP-qPCR. b Yeast one-hybrid assay of the interaction of NtCRF1 with M3-M5. Empty pGAD424: control. c ChIP-qPCR analysis of the enrichment of NtCRF1 with the four promoter regions. pNtCRF1::GFP, control. Error bars represent the standard error (SE) of three biological replicates. d Binding of recombinant NtCRF1 to M3 was outcompeted by purified NtCRF1 (2 and 4×) and biotin-labeled DNA fragments. e Top: Binding of NtCRF1 to M3 mutants was reduced; Bottom: The sequences of M3 and the three mutant motifs. f Structure of the pNtCYS-driven and mpNtCYS (mutant promoter)-driven Dual-Luc reporter gene. 35S promoter (white arrow), pNtCYS (black arrow), mpNtCYS (gray arrow), Renilla luciferase (REN), firefly luciferase (LUC) and terminator (T) are indicated. Box: The mutant motifs of mpNtCYS. g Relative reporter activity (LUC/REN) in tobacco protoplasts. The relative LUC activity was normalized to the REN activity (LUC/REN, n = 4). (Student’s t-test; ns, P > 0.05; *P < 0.05; **P < 0.01; ***P < 0.001; ****P < 0.0001). The source data of the uncropped immunoblots are provided in the Source Data file
