Extended Data Fig. 3: Time course expression analysis of ECM degradation, NOTCH, NRG1 and VEGFA pathway markers.
From: Control of cardiac jelly dynamics by NOTCH1 and NRG1 defines the building plan for trabeculation
Related to Figs. 1–3. a–m, Detailed time-course analysis of marker expression using ISH and immunofluorescence in embryonic left ventricle at E8.0, E8.5, E9.0, E9.5, E10.5, E11.5, E13.5 and E16.5 in wild-type embryos (a–e, g, i, k–m) as well as in Tie2creNotch1fl/fl (f, h) and Nrg1tm/tm (j) embryos relative to somite-matched wild-type embryos at E8.0, E8.5, E9.0 and E9.5. Data are organized according to respective ECM degradation, Notch, neuregulin-1 (NRG1) and VEGFA pathways. a–d, Analysis of ECM degradation markers Adamts1 (a), Mmp2 (b) and Hyal2 (c) by ISH and neoversican (d) by immunofluorescence. e–h, NRG1 pathway analysis of Nrg1 by RNA-scope (e, f) and pERBB2 (g, h) by immunofluorescence in wild-type (e, g) and Tie2creNotch1fl/fl (f, h) embryos. i–k, Notch pathway analysis of N1ICD (i, j) in wild-type (i) and Nrg1tm/tm (j) embryos, and DLL4 in wild-type embryos (k) by immunofluorescence. l, m, VEGFA pathway analysis of Vegfa by ISH (l) and VEGFR2 by immunofluorescence (m). In all panels showing gene/protein expression: marker expression (arrow), reduced marker expression (arrowhead), increased marker expression (thick arrow). In all immunofluorescence images: markers associated with endocardium or ECM (red); markers associated with myocardium (orange); myocardium is stained for SMA (green dulled mask); nuclei (DAPI, blue). a–m, n = 3 independent embryos were used for each genotype. Scale bars, 20 μm (E8.0), 25 μm (E8.5), 30 μm (E9.0, E9.5), 50 μm (E10.5), 60 μm (E11.5), 120 μm (E13.5) and 200 μm (E16.5).
