|
Display additional annotations [+]
|
| |
Figure 2. (A–C) Modulators of tissue compliance applied during stages of early heart development exhibit defects, including pericardial and neural edemas (A) (see arrows; scale bar, 1 mm), altered AP length (B), and increased rates of edema per clutch (C) (n = 30–35 embryos over four clutches).
(D) Compliance measured by microaspiration of HFR.
(E) Compliance at stage 22 confirms that blebbistatin and Y27632 increase and calyculin A decreases compliance (n = 11–17 embryos per treatment over three clutches).
(F) Transverse sections through HFR at stage 28 show changes in polarity (aPKC or ZO-1) within the progenitor population (red). Lower panels show the epithelial marker masked using tropomyosin expression (scale bar, 50 μm).
(G) Apical intensity after small-molecule inhibitor treatment (n = 9–13 embryos over four clutches).
(H) Representative lateral confocal sections of stage 39 tadpole hearts (scale bar, 100 μm).
(I) Cardiac anatomy after stage-specific inhibitor treatments as shown in Figure S2 (n = 5 embryos per treatment per period).
Error bars represent mean ± SEM. ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001. See also Figures S1, S2, and S3A. |
|
Display additional annotations [+]
|
| |
Figure S1. Schematic of epithelial marker apical intensity calculations. Related to Figure 1, Figure 2, Figure 5
and Figure 6.
(A) Transverse sections of stage 28 control embryo stained for tropomyosin (red) and ZO-1 (green). To calculate
apical intensity, the red and green channels are first split. The tropomyosin channel is used to segment out and create
a mask for cardiomyocytes. The ZO-1 channel has intensity normalized to background signal in the endoderm and
undergoes a small Gaussian blur (kernel = 2 pixels). (B) Applying the mask to the normalized ZO-1 channel
generates an image with only ZO-1 expression in cardiomyocytes. A 10-pixel wide linear region around along the
apical surface is created to measure apical intensity. Scale bar = 50 μm. (C) 3D projection of the ventral,
cardiomyocyte-masked, ZO-1 expression, showing apical intensity measurements at 4 different z-locations. |
