Supplementary Materials

Egg Cell–Secreted EC1 Triggers Sperm Cell Activation During Double Fertilization

Stefanie Sprunck, Svenja Rademacher, Frank Vogler, Jacqueline Gheyselinck, Ueli Grossniklaus, Thomas Dresselhaus

Materials/Methods, Supplementary Text, Tables, Figures, and/or References

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  • Materials and Methods
  • Supplementary Text
  • Figs. S1 to S10
  • Tables S1 to S3
  • Full Reference List

Images, Video, and Other Other Media

Movie S1
Merged bright field and fluorescence images of 28 serial optical sections generated by confocal laser scanning microscopy (CLSM). EC1-GFP (green signal) is secreted by the egg cell (ec) when the two sperm cells (sc1 & sc2; red fluorescent nuclei) arrive at the site of gamete interactions. Each frame of this scan represents one optical section of 0.48 μm, the z-stack size is 13.44 µm. Two individual optical sections of this scan are shown in Fig. 2F and in the inset of Fig. 2F. The scan was performed 7.5 hours after handpollinating pistils transgenic for EC1.1p:EC1-GFP with the marker line HTR10-mRFP1. The positions of the central cell (cc), the egg cell (ec) and the degenerating synergid (dsyn) are indicated. The same scan is shown in movie S2, as overlay of red and green signals only.
Movie S2
Overlay of red and green fluorescent images of the same CLSM scan shown in movie S1. Secretion of EC1-GFP (green signal) can be observed when the two sperm cells (sc1 & sc2; red fluorescent nuclei) arrive at the site of gamete interactions. Each frame of this scan represents one optical section of 0.48 µm, the z-stack size is 13.44 μm. Two individual optical sections of this scan are shown in Fig. 2G and in the inset of Fig. 2G. The scan was performed 7.5 hours after hand-pollinating pistils transgenic for EC1.1p:EC1-GFP with the marker line HTR10-mRFP1. The positions of the central cell (cc), the egg cell (ec) and the degenerating synergid (dsyn) are indicated. The same scan is shown in movie S1 as merged bright field and fluorescence images.