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. 2020 May 12;31(6):107627. doi: 10.1016/j.celrep.2020.107627

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© 2020 The Author(s)

This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).

PMC Copyright notice

Functional HSCs Are Localized in 1- to 2-Cell IAHC Cells

(A–D) Sections through dorsal aorta of E11 (43 somite pairs, sp) Gata2Venus (green in A and B) and Ly6aGFP (green in C) and of E10.5 (36sp) Csf1rGFP (MacGreen; green in D) embryos. Nuclei are indicated in blue (DAPI), endothelial and IAHC cells are indicated in red (CD34) and CD27⁺ cells are indicated in white. v, ventral, d, dorsal.

(A) Arrowhead indicates small cluster containing two G2⁺CD27⁺ cells attached to one G2⁺CD27⁻ cell. Arrow indicates a single rounded/bulging G2⁺CD27⁺ cell.

(B) In (Bi), ventral cluster with two G2^medCD27⁺ cells (one bulging out of endothelium). (Bii) Ventral two-cell cluster (G2^medCD27⁺ and G2^loCD27⁻ cells). (Biii) Single rounded bulging G2⁺CD27⁺ cell on dorsal side of aorta.

(C) In (Ci), single Ly6aGFP⁺CD27⁺ cell (arrowhead) at apex of bulging Ly6aGFP⁺ cell within ventral endothelium. A neighboring GFP⁺ cell in the ventral endothelium is CD27⁻ (arrow). (Cii) Two round CD27⁺ cells (one is Ly6aGFP^lo, indicated by arrowhead) adjacent to each other and attached to interspersed Ly6aGFP⁺ endothelium. (Ciii) Three CD27⁺ cells (indicated by arrowheads; one is Ly6aGFP⁺) in very close association with ventral endothelium. White dashed boxes indicate magnified areas.

(D) One CD27⁺CD34⁺ cell on dorsal endothelial wall closely associated with a macrophage. White dashed box indicates area for which MacGreen and CD27 expression is shown.

Scale bars, 20 μm.