Extended Data Fig. 1: Generation of an inducible Npr3CreERT2 mouse line for fate mapping of perinatal ventricular endocardial cells.
From: Perinatal angiogenesis from pre-existing coronary vessels via DLL4–NOTCH1 signalling
a, RNAscope ISH and immunofluorescence images showing Npr3 transcripts (red) and EMCN (green) in ventricular endocardial cells of P0 hearts. The images are representative of three individual hearts. tb, trabecular myocardium; com, compact myocardium; pm, papillary muscle. b, Schematic showing the strategy for generating a tamoxifen-inducible Npr3CreERT2 mouse line for fate mapping of perinatal ventricular endocardial cells. c, Co-immunofluorescence images showing expression of estrogen receptor (ER, indicative of CreERT2, red) and EMCN (green) in ventricular endocardium (en), but not coronary plexus, of E17.5 Npr3CreERT2 heart. The images are representative of four individual hearts. d, Co-immunofluorescence showing comparable expression level and pattern of NPR3 between wild-type and Npr3CreERT2 neonatal heart at P0. Some small red peppered background signals are associated with the NRP3 antibody staining. Arrows indicate coronary artery. The images are representative of 4 individual hearts. e, RT-qPCR analysis indicating comparable expression of Npr3 transcripts between wild type and Npr3CreERT2 perinatal heart at E17.5 (n=5 hearts for each group) and P0 (n=4 hearts for each group). Data are presented as mean ± SEM and two-tailed unpaired Student’s t-test. All samples were derived from biologically independent experiments. Scale bar: 500 µm in a, c, d.
