Chick cardiac cells before and during C looping exhibit dominant CW rotation in vitro. Chick cardiac cells before and during C looping exhibit dominant.

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Chick cardiac cells before and during C looping exhibit dominant CW rotation in vitro. Chick cardiac cells before and during C looping exhibit dominant CW rotation in vitro. (A) Confocal images of the chicken embryonic heart between HH9 and HH11. Blue arrow indicates the rightward direction of cardiac rotation at HH10. Red, F-actin (Phalloidin); green, anti-ZO1; blue, nucleus (Hoechst). (Scale bar: 200 µm.) A, anterior; E, endocardium; L, left; M, myocardium; OV, omphalomesenteric veins; P, posterior; R, right; and SPL, splanchnopleure. (B) Overview of culture of cardiac cells at the interface of a bilayered Matrigel system. Cells were isolated from the chicken embryonic heart tube during the looping stages, followed by a brief culture in the bilayered Matrigel system before analyzing the directionality of rotation using time-lapse microscopy. (C) MF20 antibody staining showing the abundance of myocardial cells in Matrigel during 3D culture. (Scale bar: 150 µm.) (D) Percentage of cardiac cells isolated from chicken embryos between HH9 and HH11 undergoing CW, CCW, no rotation, complex rotation, and rotation. (E) Table denoting total number of cells from different experiments undergoing CW, CCW, no rotation, complex rotation, and rotation. Boldface type indicates a dominant bias. *P < 0.05, ***P < 0.001; ns, nonsignificant. Poulomi Ray et al. PNAS doi:10.1073/pnas.1808052115 ©2018 by National Academy of Sciences