Extended Data Fig. 9: Ectopic expression of truncated versions of ECS1 and ECS2 or the subtilisin-like protease SBT4.13 in synergid cells have no significant influence on LURE1.2 protein levels and pollen tube attraction rate.
From: Fertilized egg cells secrete endopeptidases to avoid polytubey
a, Ectopic expressions of truncated versions of ECS1 and ECS2 (TECS1/2) in synergid cells. b, Proportions of ovules attracting pollen tubes observed in WT plants and those ectopically expressing TECS1/2 in synergid cells at 6 HAP (n = 50). c, Immunofluorescence revealed that LURE1 levels were comparable in WT and ovules ectopically expressing TECS1/2 in synergid cells using the DD31 promoter. d, Quantification of LURE1 fluorescence intensity in WT ovules and those ectopically expressing truncated TECS1/2 in synergid cells (n = 10). e, Ectopic expressions of the egg cell expressed subtilisin-like protease SBT4.1327 as a GFP fusion protein in synergid cells. SBT4.13–GFP is secreted to the filiform apparatus. f, Proportions of ovules attracting pollen tubes observed in WT plants and those ectopically expressing SBT4.13 in synergid cells at 6 HAP (n = 50). g, Immunofluorescence revealed that LURE1 levels were comparable in WT and ovules ectopically expressing SBT4.13 in synergid cells. h, Quantification of LURE1 fluorescence intensity in WT ovules and those ectopically expressing SBT4.13 (n = 10). Data in b, f represent the mean ± s.d. Same letters in b, f indicate lack of significant differences according to the Tukey–Kramer multiple comparison test (one-way ANOVA between groups; P = 0.96, F = 0.04 in b left plane; P = 0.85, F = 0.17 in b right plane; P = 0.85, F = 0.17 in f). P < 0.05 was considered as significant. Data in d, h 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. Two-tailed Student’s t-test was used for statistical test in d, h. ns, no significant differences. sy, synergid cell. Scale bars, 20 μm.
