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. 2021 May 10;118(20):e2021814118. doi: 10.1073/pnas.2021814118

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Copyright © 2021 the Author(s). Published by PNAS.

This open access article is distributed under Creative Commons Attribution-NonCommercial-NoDerivatives License 4.0 (CC BY-NC-ND).

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Fig. 1. — CYK-1/Formin is a determinant of actomyosin flow chirality. (A) Schematic of a C. elegans zygote during anteroposterior polarization. Gray: region used to obtain the flow velocity profiles. Blue areas: regions of interest used to calculate the mean flow speed, chiral velocity, and chiral ratio. (B) Time-averaged flow field overlaid on an image of cortical NMY-2::GFP of a control and a cyk-1(lf) mutant embryo. Velocity vectors are color coded for their angle with the anteroposterior axis. (Scale bar, 10 $μ$ m.) Velocity scale arrow, 20 $μ$ m/min. (C) Mean y velocity in 18 bins along the anteroposterior axis averaged over embryos for control (gray) and for cyk-1(lf) (blue). Light blue areas: bins 3 to 6 and 13 to 16 corresponding to the anterior and posterior regions of interest, respectively. Error bars, SEM. (D) Mean speed per embryo defined as $({< | v | >}_{a} + < | v {| >}_{p}) / 2$ , where ${< | v | >}_{a}$ and ${< | v | >}_{p}$ are the spatial averages in the anterior and posterior regions of interest, respectively. (E) Mean chiral velocity, $v_{c}$ , per embryo defined as $< {v_{y} >}_{p} - < {v_{y} >}_{a}$ , where $< {v_{y} >}_{a}$ and $< {v_{y} >}_{p}$ are the spatially averaged y velocities in the anterior and posterior regions of interest, respectively. (F) Mean chiral ratio per embryo, defined as $\frac{- v_{c}}{< | v {| >}_{a} + < | v {| >}_{p}}$ . (G) Domain overview of CYK-1/Formin (top) and the constructs generated in this study (middle and bottom). DID, Dia Inhibitory Domain; DD, Dimerization Domain; CC, Coiled Coil; FH, Formin Homology; DAD, Diaphanous Autoregulatory Domain; PH, Pleckstrin Homology; LOV2, Light-Oxygen-Voltage 2 domain. (H) Time-averaged flow field overlaid on an image of cortical Lifeact-mKate2 derived from control (ph-gfp-lov2) injection and ca-cyk-1/Formin injection. (I) Mean y-velocity profile in embryos derived from control injection (gray) and from ca-cyk-1/Formin injection (blue). (J–L) Mean flow speed (J), chiral velocity (K), and chiral ratio (L) in embryos derived from control injection and from ca-cyk-1/Formin injection. Blue in D–F and J–L depicts the mean over embryos with 95 $%$ confidence interval. Significance testing: *P $\leq$ 0.05; **P $\leq$ 0.01; ***P $\leq$ 0.001 (Wilcoxon rank sum test). n indicates the number of embryos.