Fig. 7: OsSRF8 recruits OsDAF1 to regulate annulus formation.
From: OsSRF8 interacts with OsINP1 and OsDAF1 to regulate pollen aperture formation in rice
a Y2H assay verifies the interaction of OsSRF8 and OsDAF1. OsDAF1 (N) and OsDAF1 (C) represent the N-terminal domain (amino acids 24-331) and C-terminal domain (amino acids 355-695) of the OsDAF1 protein, respectively. b Split-luciferase assay testing the interaction between OsSRF8 and OsDAF1 in the tobacco (Nicotiana benthamiana) leaves. c Interactions between OsSRF8 and OsDAF1 detected using BiFC assay. OsSRF8 and OsDAF1respectively were fused to the N- and C-terminal parts of YFP and co-transformed into tobacco leaves. Scale bars, 50 μm. d Co-immunoprecipitation experiments in the rice protoplasts (left) and tobacco leaves (right). α-Flag IP indicates immunoprecipitation with anti-Flag antibody beads. Experiments in (a–d) were repeated three times, with similar results. Confocal images and illustrations of OsDAF1-eYFP signal in tetrads of WT (e) and Ossrf8 mutant (f) backgrounds. Scale bars, 10 μm. g Confocal optical sections and schematic diagrams of tetrads expressing OsSRF8-eYFP protein in the Osdaf1 background. Five independent T1 lines were imaged, with similar results. Scale bars, 10 μm. The graphs on the right represent the fluorescence values after quantification and the white dashed arrows indicate the direction of fluorescence measurement.
