Tagging RHGF-1 with tagRFP does not perturb its function and shows that following oocyte entry RHGF-1 relocalizes from the cytoplasm to the basal side. (A and B) Comparison of dwell time and successful embryo transits in wild-type worms versus worm in which genomic RHGF-1 was fused with tagRFP. N ≥ 18. Error bars are mean ± SEM. Statistical comparison for A was performed by two-tailed unpaired t test. Statistical comparison for B was performed by chi-square-Fisher’s exact test. (C) Fluorescence intensity of RHGF-1::tagRFP treated with rhgf-1(RNAi), normalized to control fluorescence intensity. N ≥ 19. Bars are mean ± SEM. Statistical comparison was performed by two-tailed unpaired t test with Welch’s correction. (D) Confocal images from a time-lapse movie of a spermatheca expressing RHGF-1::tagRFP and β-Integrin/PAT-3::GFP as a basal membrane marker. Scale bar, 10 µm. Full movie in Video 6. (E) Representative images demonstrating how we measured the intensity of SPV-1::GFP and RHGF-1::tagRFP in the cytoplasm (yellow) and edge (green), respectively. Region in black box enlarged on right. Scale bars: whole spermatheca, 20 µm; enlargement, 2 µm. Stars designate statistical significance (**** P <0.0001, ** P <0.01, ns P >0.05). N, number of samples analyzed.