Fig. 1: NOTCH1 and JAG1 are co-expressed in T2-HSCs.
From: Cis inhibition of NOTCH1 through JAGGED1 sustains embryonic hematopoietic stem cell fate
A scheme of the IAHC formation in the mouse embryo. GFI1 + HE cells undergo EHT and form IAHC within the dorsal aorta. Within these clusters, the GFI1 + IAHC fraction contains all HSC activity. The HSC population can be further restricted by including CD45 as a marker of T2-HSCs (T2-like HSC). B Representative Flow cytometry plot of AGM lysates stained for CD31 and cKIT. The gate for GFI1+ cells within each population is superimposed onto the plot. C Representative Flow Cytometry plots of HE with superimposed gates for indicated notch signaling molecule. D Quantification of NOTCH1, JAG1 and DLL4 positive cells within the HE population at E10.5 and E11.5. 2-5 AGMs were pooled for each data point in 4 independent experiments (E10.5 n = 18 embryos, E11.5 n = 22 embryos). Statistical significance was calculated with two-tailed t-tests. Vertical error bars indicate the mean and standard deviation values. Source data are provided as a source data file. E Exemplary Flow Cytometry plots of GFI1 + IAHC with superimposed gates for indicated notch signaling molecule. F Quantification of NOTCH1, JAG1 and DLL4 positive cells within the GFI1 + IAHC population at E10, E10.5 and E11.5. 2-18 AGMs were pooled for each data point in 4 independent experiments (E10 (27-33 s) n = 32 embryos, E10.5 n = 18 embryos, E11.5 n = 22 embryos). Statistical significance was calculated with two-tailed t-tests. Vertical error bars indicate the mean and standard deviation values. Source data are provided as a source data file. G Representative Flow Cytometry plots showing JAG1 and NOTCH1 levels within GFI1 + IAHC (CD31+cKIT+GFI1 + ) sub gated for CD45 at E10.25 and E11.5. NOTCH1-JAG1 double positive cells in the CD45+ fraction is highlighted in red. H Line chart of Notch1/JAG1 double positive cells within CD45 + /- IAHC (CD31+cKIT + ) between E10 and E11.5. 2-5 AGMs were pooled for each data point in E10 (27–28 s, n = 8 embryos, E10.25 (31–33 s, n = 24 embryos, E10.5 (34–36 s, n = 18 embryos and E11.5 (36-46, n = 22 embryos) 4 independent experiments for each time point. Statistical significance was calculated with two-tailed t-tests. Vertical error bars indicate the mean and standard deviation values. Source data are provided as a source data file. (s: number of somites).
