Figure S1.
LIN-5 regulates timely furrow induction independent of its role in spindle elongation. (A) Schematics of the cell cycle progression, indicating various stages: NEBD (i), anaphase onset (ii), and furrow involution (iii). (B) Quantification of the cell cycle interval between the above-indicated stages (i), (ii), and (iii) from the confocal time-lapse movies of embryos coexpressing GFP-tagged PLCδ1-PH (membrane marker), GFP-TBB-2 (microtubule marker), mCherry-H2B (chromatin marker), and mCherry-γ-tubulin (centrosomal marker), referred to as GFPMem./Tub.;mCherryChr./Cent. in control, zen-4(RNAi), lin-5(RNAi), gpr-1/2(RNAi) embryos, as indicated. The time interval between stages (i) and (ii) in control, zen-4(RNAi), lin-5(RNAi), and gpr-1/2(RNAi) embryos is nonsignificant (ns). However, there is a significant increase in a time interval between stages (ii) and (iii) of ∼60 s in lin-5(RNAi) and ∼70 s in gpr-1/2(RNAi) when compared to control and zen-4(RNAi). Consequently, a similar delay persists from the stage (i) to (iii). n is the number of embryos analyzed. Error bars are the SEM; ns, P > 0.05; ***P < 0.001, as determined by two-tailed Student’s t test. (C–F) Representative pseudokymographs of the division plane (shown on the top right) acquired by time-lapse confocal microscopy of control embryos coexpressing GFPMem./Tub.;mCherryChr./Cent.. (G) These embryos were either left untreated (C), depleted for LIN-5 (D), depleted for GPR-1/2 (E), depleted for ZEN-4 (F), or codepleted for LIN-5 and ZEN-4 (G), as indicated. Please note re-use of the lin-5(RNAi) pseudokymograph in Fig. 1 D. Control, n = 17; lin-5(RNAi), n = 11, gpr-1/2(RNAi), n = 8, zen-4(RNAi), n = 10, lin-5(RNAi); zen-4(RNAi), n = 10. For comparative analysis, only GFPMem./Tub. localization is shown. Pseudokymographs start 200 s after NEBD, which was deduced by the entry of tubulin in the pronuclei. n is the number of embryos analyzed. The scale bar is 5 µm. (H–M) Representative images from the time-lapse confocal microscopy of embryo coexpressing GFPMem./Tub.;mCherryChr./Cent. in control (H) and in various RNAi/mutant conditions, as indicated (I–M): control (n = 12); lin-5(RNAi) (n = 19); hcp-4(RNAi) (n = 13); lin-5(RNAi); hcp-4(RNAi) (n = 11); spd-1(ts) (n = 13); lin-5(RNAi) in spd-1(ts) (n = 12). n is the number of embryos analyzed. In this and subsequent one-cell embryo images, the posterior is to the right. Time is in seconds (s) w.r.t. NEBD. The scale bar is 10 µm.

LIN-5 regulates timely furrow induction independent of its role in spindle elongation. (A) Schematics of the cell cycle progression, indicating various stages: NEBD (i), anaphase onset (ii), and furrow involution (iii). (B) Quantification of the cell cycle interval between the above-indicated stages (i), (ii), and (iii) from the confocal time-lapse movies of embryos coexpressing GFP-tagged PLCδ1-PH (membrane marker), GFP-TBB-2 (microtubule marker), mCherry-H2B (chromatin marker), and mCherry-γ-tubulin (centrosomal marker), referred to as GFPMem./Tub.;mCherryChr./Cent. in control, zen-4(RNAi), lin-5(RNAi), gpr-1/2(RNAi) embryos, as indicated. The time interval between stages (i) and (ii) in control, zen-4(RNAi), lin-5(RNAi), and gpr-1/2(RNAi) embryos is nonsignificant (ns). However, there is a significant increase in a time interval between stages (ii) and (iii) of ∼60 s in lin-5(RNAi) and ∼70 s in gpr-1/2(RNAi) when compared to control and zen-4(RNAi). Consequently, a similar delay persists from the stage (i) to (iii). n is the number of embryos analyzed. Error bars are the SEM; ns, P > 0.05; ***P < 0.001, as determined by two-tailed Student’s t test. (C–F) Representative pseudokymographs of the division plane (shown on the top right) acquired by time-lapse confocal microscopy of control embryos coexpressing GFPMem./Tub.;mCherryChr./Cent.. (G) These embryos were either left untreated (C), depleted for LIN-5 (D), depleted for GPR-1/2 (E), depleted for ZEN-4 (F), or codepleted for LIN-5 and ZEN-4 (G), as indicated. Please note re-use of the lin-5(RNAi) pseudokymograph in Fig. 1 D. Control, n = 17; lin-5(RNAi), n = 11, gpr-1/2(RNAi), n = 8, zen-4(RNAi), n = 10, lin-5(RNAi); zen-4(RNAi), n = 10. For comparative analysis, only GFPMem./Tub. localization is shown. Pseudokymographs start 200 s after NEBD, which was deduced by the entry of tubulin in the pronuclei. n is the number of embryos analyzed. The scale bar is 5 µm. (H–M) Representative images from the time-lapse confocal microscopy of embryo coexpressing GFPMem./Tub.;mCherryChr./Cent. in control (H) and in various RNAi/mutant conditions, as indicated (I–M): control (n = 12); lin-5(RNAi) (n = 19); hcp-4(RNAi) (n = 13); lin-5(RNAi); hcp-4(RNAi) (n = 11); spd-1(ts) (n = 13); lin-5(RNAi) in spd-1(ts) (n = 12). n is the number of embryos analyzed. In this and subsequent one-cell embryo images, the posterior is to the right. Time is in seconds (s) w.r.t. NEBD. The scale bar is 10 µm.

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