Extended Data Fig. 7: ECS1 and ECS2 efficiently cleave LURE1 substrates.
From: Fertilized egg cells secrete endopeptidases to avoid polytubey
a, Localization and protein level of LURE1.2–GFP before pollination and in fertilized ovules of WT and ecs1 ecs2 mutant pistils, respectively. Pistils were pollinated with pollen expressing HTR10–mRFP in sperm cells. Ovules were collected from pistils at 0 and 10 HAP. ecs1 ecs2 mutation resulted in the accumulation of LURE1.2 after fertilization. b, Quantification of green fluorescence intensity in ovules from WT and ecs1 ecs2 pistils (n = 101). Data for fluorescence intensity are presented in box-and-whisker plots. Bottom and top of the box, 25th and 75th percentiles; centre line, 50th percentile; whiskers, minimum and maximum data. ** indicates statistically significant difference between WT and mutant ovules (two-tailed Student’s t-test; P < 0.01). c, In vitro pollen tube attraction assay with gelatin beads containing ECS1- and ECS2-digested LURE1.2, respectively. Beads (*) were prepared using 1 μM LURE1.2 alone and in combination with 1 μM ECS1 and ECS2, respectively, and placed close to growing pollen tube tips (0 min) and observed for 60 min. Pollen tube attraction activity was lost when beads contained both, LURE1.2 and ECS endopeptidases. n, nucleus; pt, pollen tube; sy, synergid cell; zy, zygote. Scale bars are 10 μm (a) and 50 μm (c).
