Figure S2.
In contrast with RHGF-1, UNC-73 does not appear to regulate RHO-1 in the spermatheca. (A) Snapshots at indicated times from movies of spermatheca-specific RHO-1 biosensor (AHPH::GFP) during ovulation and embryo transit in control, and an rhgf-1(RNAi) spermatheca that trapped the embryo for the duration of the movie. Scale bar, 20 µm. Full movies are in Video 4. (B) Quantification of RHO-1 biosensor fluorescence intensity during ovulation and embryo transit in three controls and three rhgf-1(RNAi) worms that trapped the embryo. (C) Live imaging of spermatheca-specific RHO-1 biosensor (AHPH::GFP) during embryo transit through spermatheca in control and unc-73(RNAi). Scale bar, 20 µm. Full movies are in Video 5. (D) Quantification of RHO-1 biosensor fluorescence intensity during ovulation and embryo transit in control and unc-73(RNAi) worms. 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 = 6. (E) Dwell time in rhgf-1(ok880) spermathecae treated with spermatheca-specific unc-73(RNAi). N ≥ 7. Bars are mean ± SEM. Statistical comparison was performed by two-tailed unpaired t test with Welch’s correction. (F) Successful embryos transit through the spermathecae of rhgf-1(ok880) treated with spermatheca-specific unc-73(RNAi). N ≥ 19. Statistical comparison was performed by chi-square test. (G) Fluorescence intensity of UNC-73::GFP treated with unc-73(RNAi), normalize to control fluorescence intensity. N ≥ 27. Bars are mean ± SEM. Statistical comparison was performed by two-tailed unpaired t test with Welch’s correction. Stars designate statistical significance (**** P <0.0001, ns P >0.05). N, number of samples analyzed.

In contrast with RHGF-1, UNC-73 does not appear to regulate RHO-1 in the spermatheca. (A) Snapshots at indicated times from movies of spermatheca-specific RHO-1 biosensor (AHPH::GFP) during ovulation and embryo transit in control, and an rhgf-1(RNAi) spermatheca that trapped the embryo for the duration of the movie. Scale bar, 20 µm. Full movies are in Video 4. (B) Quantification of RHO-1 biosensor fluorescence intensity during ovulation and embryo transit in three controls and three rhgf-1(RNAi) worms that trapped the embryo. (C) Live imaging of spermatheca-specific RHO-1 biosensor (AHPH::GFP) during embryo transit through spermatheca in control and unc-73(RNAi). Scale bar, 20 µm. Full movies are in Video 5. (D) Quantification of RHO-1 biosensor fluorescence intensity during ovulation and embryo transit in control and unc-73(RNAi) worms. 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 = 6. (E) Dwell time in rhgf-1(ok880) spermathecae treated with spermatheca-specific unc-73(RNAi). N ≥ 7. Bars are mean ± SEM. Statistical comparison was performed by two-tailed unpaired t test with Welch’s correction. (F) Successful embryos transit through the spermathecae of rhgf-1(ok880) treated with spermatheca-specific unc-73(RNAi). N ≥ 19. Statistical comparison was performed by chi-square test. (G) Fluorescence intensity of UNC-73::GFP treated with unc-73(RNAi), normalize to control fluorescence intensity. N ≥ 27. Bars are mean ± SEM. Statistical comparison was performed by two-tailed unpaired t test with Welch’s correction. Stars designate statistical significance (**** P <0.0001, ns P >0.05). N, number of samples analyzed.

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