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Display additional annotations [+]
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Figure S7. Microsurgical manipulations to visualize HPCs in Xenopus. Related to Experimental Procedures.
(A) Schematic of microsurgical manipulations to visualize HPC migration. (B) Microsurgical isolates at 1 h and 24
h. (C) Ventral windowed embryo at 1 h and 24 h. (D) Lateral windowed embryo at 1 h and 24 h. Yellow arrows
mark location of beating heart. Scale bar = 500 μm. (E) Stage 33 embryo and equivalent stage microsurgical isolate
stained for troponin T (red), fibrillin (white) and nuclei (green). Scale bar = 50 μm. |
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Display additional annotations [+]
Gene |
Clone |
Species |
Stages |
Anatomy |
prkci
|
|
laevis
|
NF stage 28
|
ectoderm
,
mesoderm
,
cardiac myocyte
,
primary heart field
|
tpm1
|
|
laevis
|
NF stage 28
|
mesoderm
,
cardiac myocyte
,
primary heart field
|
tjp1
|
|
laevis
|
NF stage 28
|
ectoderm
,
mesoderm
,
endoderm
,
cardiac myocyte
,
primary heart field
|
fbn1
|
|
laevis
|
NF stage 39
|
heart
,
endocardium
|
|
|
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. |