Figure 3.
RHGF-1 is expressed in the spermatheca and localizes to the basal side following oocyte entry and parallel to RHO-1 activation. (A) RHGF-1::tagRFP expression pattern in the adult hermaphrodite by confocal image (left) and merged with DIC image (right). Arrows point to an occupied spermatheca and arrowheads point to an empty spermatheca. Image is a composite of multiple images stitched in Fiji. Scale bar, 50 µm. (B) Representative confocal images showing the subcellular localization of both RHGF-1::tagRFP and SPV-1::GFP in the same spermatheca from ovulation to the completion of embryo exit. White boxes mark regions enlarged below the images. The graphs show the intensity line profile along the marked lines from the apical side of the cell (“A”) to the basal side (“B”). Dashed lines mark the edges of spermathecal cells. Scale bars: whole spermatheca, 10 µm; enlargement, 2 µm. Full movie in Video 7. (C) Quantification of both RHGF-1::tagRFP and SPV-1::GFP recruitment to the cell edge during ovulation. The x-axis (time) was normalized according to distinct events so that all traces begin at the time when the distal valve closes behind the ovulating oocyte (“Oocyte entry complete”) are aligned at the time the sp-ut starts to open (“Embryo exit initiation”) and aligned again when the sp-ut valve closes behind the exiting embryo (“Embryo exit complete”). N = 3. (D) Representative confocal images of RHGF-1 subcellular localization and RHO-1 biosensor, in the same spermatheca, during oocyte ovulation and embryo transit. White boxes mark regions enlarged below the images. Dashed lines mark the edges of spermathecal cells. Scale bars: whole spermatheca, 20 µm; enlargement, 2 µm. Full movie in Video 8. (E) Quantification of RHGF-1::tagRFP recruitment to the cell edge, and RHO-1 biosensor intensity during ovulation and embryo exit. The x-axis (time) was normalized as in C. N = 6. N, number of samples analyzed.

RHGF-1 is expressed in the spermatheca and localizes to the basal side following oocyte entry and parallel to RHO-1 activation. (A) RHGF-1::tagRFP expression pattern in the adult hermaphrodite by confocal image (left) and merged with DIC image (right). Arrows point to an occupied spermatheca and arrowheads point to an empty spermatheca. Image is a composite of multiple images stitched in Fiji. Scale bar, 50 µm. (B) Representative confocal images showing the subcellular localization of both RHGF-1::tagRFP and SPV-1::GFP in the same spermatheca from ovulation to the completion of embryo exit. White boxes mark regions enlarged below the images. The graphs show the intensity line profile along the marked lines from the apical side of the cell (“A”) to the basal side (“B”). Dashed lines mark the edges of spermathecal cells. Scale bars: whole spermatheca, 10 µm; enlargement, 2 µm. Full movie in Video 7. (C) Quantification of both RHGF-1::tagRFP and SPV-1::GFP recruitment to the cell edge during ovulation. The x-axis (time) was normalized according to distinct events so that all traces begin at the time when the distal valve closes behind the ovulating oocyte (“Oocyte entry complete”) are aligned at the time the sp-ut starts to open (“Embryo exit initiation”) and aligned again when the sp-ut valve closes behind the exiting embryo (“Embryo exit complete”). N = 3. (D) Representative confocal images of RHGF-1 subcellular localization and RHO-1 biosensor, in the same spermatheca, during oocyte ovulation and embryo transit. White boxes mark regions enlarged below the images. Dashed lines mark the edges of spermathecal cells. Scale bars: whole spermatheca, 20 µm; enlargement, 2 µm. Full movie in Video 8. (E) Quantification of RHGF-1::tagRFP recruitment to the cell edge, and RHO-1 biosensor intensity during ovulation and embryo exit. The x-axis (time) was normalized as in C. N = 6. N, number of samples analyzed.

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