Fig. 5: Molecular interaction between Sh1 and SHAT1-5 and cellular mechanism for shattering.

a Schematic diagram of SHAT1-5 promoter segments used in Y1H assay. b Results of Y1H assays with SHAT1-5 promoter segments. Diagram illustrating the results of Y1H assays. From left to right, S1 to S5. c Schematic diagram of the SHAT1-5 promoter S2 segments fragmentation (60 bp for each). d Results of Y1H assays with SHAT1-5 promoter S2 segments fragmentation. Diagram illustrating the results of Y1H assays. For the plate have seven groups, both 3 and 5 indicate S2-1, S2-3, S2-5, S2-7, S2-9, and S2-11 segments, and both 4 and 6 indicate S2-2, S2-4, S2-6, S2-8, and S2-12 segments, respectively. e EMSA detection of Sh1 binding to S2-5, S2-6, S2-11 and S2-12 segments of SHAT1-5 promoter. EMSA of 3’-biotin-labeled dsDNA probes with the purified Sh1 protein. The presence (+) or absence (–) of specific probes is marked. The concentration of the cold probe was 1 μM (100x), 2 μM (200x), while that of the biotinylated probe was 10 nM. Water was added in place of Sh1 protein as a control. f Regulation of Sh1 to SHAT1-5 using the dual luciferase assay. Relative LUC/REN activities after infiltration for 3 days. g The expression of SHAT1-5 in R3 pods of Sh1 transgenic plants. h Cross section analysis of ventral suture of Sh1/sh1 and Williams 82. The lignified fiber cap cell is indicated by arrows. n = 6 biological samples for f, and n = 3 biological samples for g, respectively. For f, g, data are presented as mean values ± SD, and the statistical significance is determined by a two-sided t test. ***, **, * indicate P < 0.001, 0.01, and 0.05, respectively. For e, h, three independent replicates are performed, and a representative result is shown. Source data are provided as a Source Data file.