Extended Data Fig. 6: Opposing gradients of gene expression in inner versus outer preexisting coronary vessels, and generation of Dll4+ lineage tracing mice.
From: Perinatal angiogenesis from pre-existing coronary vessels via DLL4–NOTCH1 signalling
a,b, Multicolor RNAscope for Dll4 (red) or Vegfr3 (red) with Aplnr (green) showing that Dll4 or Vegfr3 expressing angiogenic cells are mainly localized to the inner myocardium, whereas Aplnr-expressing cells are present in the outer myocardium at E16.5. c,d, RNAscope for Dll4, Vegfr3 or Aplnr (red) and co-immunofluorescence for EMCN (green) showing that Dll4 or Vegfr3 expressing angiogenic cells are mainly localized in the inner myocardium expressing low-to-negative levels of EMCN, whilst Aplnr-expressing cells are present in the outer myocardium expressing high levels of EMCN in the perinatal heart between E17.5 and P0. ca: coronary artery. Scale bar: 500 µm. e,f, Strategy for generating an inducible Dll4CreERT lineage-tracing mouse line and lineage tracing workflow. All samples were derived from biologically independent experiments. The images in a, b, c, and d are representative of three hearts in each experiment. Scale bar: 500 µm in a-d.
