NMY-2 membrane stripping by LIN-5 is not a critical determinant for cleavage furrow induction. (A and B) Schematic of the method to quantify the line scan intensity of GFP-NMY-2 (A) and its quantification along the A-P-A axis (B), as indicated. Black asterisks in A and B indicate NMY-2 clearance at the posterior cortical membrane near the posterior furrow involution region. (C and D) Representative images from the time-lapse confocal microscopy of embryos coexpressing GFP^Mem. and mCherry^Tub. in zyg-9(RNAi) (C), and zyg-9(RNAi) and lin-5(RNAi) in the lin-5(ts) background (D). White arrowheads indicate furrow, and yellow asterisks indicate the dissolution of the furrow in zyg-9(RNAi) embryos inhibited for LIN-5. Note the re-use of the zyg-9(RNAi) embryo image shown in Fig. S4C for Fig. 3 K. The scale bar is 10 µm. (E and F) Schematic of the method used to measure the pole-to-pole distance at the time of furrow initiation in embryos coexpressing GFP^Mem. and mCherry^Tub. (E). Quantification of pole–pole distance at the time of furrow onset/involution (F) in zyg-9(RNAi) and in zyg-9(RNAi) and lin-5(RNAi) in the lin-5(ts) background, as indicated. Each dot represents one embryo, and n is the number of embryos analyzed. Mean is represented in the black line. Error bars, SEM. ns, P > 0.05; ***P < 0.001, as determined by two-tailed Student’s t test. (G) Representative images from the time-lapse confocal microscopy of embryos endogenously expressing LIN-5-mNG in zyg-9(RNAi); asterisks indicate LIN-mNG signal at the posterior cortex, and white arrowheads indicate the presence of cortical LIN-5-mNG at the invaginating furrow. The scale bar is 10 μm. Boxed regions are the magnified view of the corresponding inset. The scale bar for the inset is 5 μm.