Figure S4.
sma-1(ru18) and sma-1(RNAi) rings constrict at a normal rate and with normal levels of actin and myosin, and both perturbations exacerbate ruptures of cyk-1(RNAi) rings without affecting F-actin levels. (A) Images of control and sma-1(ru18) ABa cells co-expressing LifeAct::GFP, NMY-2::mKate2 and mCherry::HIS-58. (B) Ring constriction rate of ABa/p rings (mean ±95% CI). n is the number of contractile rings analyzed (n = 11 or n = 10 in control, n = 18 or n = 16 in cyk-1(RNAi), n = 11 in sma-1(ru18), n = 10 in sma-1(RNAi), n = 35 in cyk-1(RNAi);sma-1(ru18), and n = 19 in cyk-1(RNAi);sma-1(RNAi)). (C) Instantaneous ring perimeter change (10 s interval) versus ring perimeter in ABa/p rings throughout constriction. 16 contractile rings were analyzed per condition. (D) Ring closure profiles in ABa/p cells. The profile mean (±95% CI) is shown for control and cyk-1(RNAi). Four individual profiles of cyk-1(RNAi);sma-1(ru18) are shown in blue, yellow, pink, and purple. Values of perimeters of cyk-1(RNAi);sma-1(ru18) rings at the time when large ruptures occurred, and constriction rates measured immediately after rupture, are shown in corresponding colors. n is the number of contractile rings analyzed (n = 11 in control and n = 18 in cyk-1(RNAi)). (E) Images of ABa rings from embryos co-expressing LifeAct::GFP, NMY-2::mKate2, and mCherry::HIS-58. (F) Normalized mean fluorescence intensity of LifeAct::GFP (±95% CI) in the peripheral arc of ABa rings. n is the number of contractile rings analyzed (n = 6 in control, n = 6 in cyk-1(RNAi), n = 7 in sma-1(ru18), and n = 9 in cyk-1(RNAi);sma-1(ru18)). (G) Images of dividing ABa cells co-expressing SMA-1::GFP (split GFP), NMY-2::mKate2, and mCherry::HIS-58. In B and F, statistical significance was determined using unpaired one-way ANOVA followed by Bonferroni’s multiple comparison test. ****P < 0.0001; ns, not significant, P ≥ 0.05. Scale bars in A, E, and G, 5 µm.