Transcription
Cell Stem Cell, volume 9Supplemental InformationErythroid/Myeloid Progenitorsand Hematopoietic Stem Cells Originatefrom Distinct Populations of Endothelial CellsMichael J. Chen, Yan Li, Maria Elena De Obaldia, Qi Yang, Amanda D. Yzaguirre,Tomoko Yamada-Inagawa, Chris S. Vink, Avinash Bhandoola, Elaine Dzierzak, and NancyA. Speck
Figure S1 (related to Figure 3). The Ly6a-GFP/Cbfb transgene fails to restore fetal viability.A. Phenotype of Cbfb / , Cbfb-/-; Tg(Ly6a-GFP/Cbfb), and Cbfb-/-; Tg(Tek-GFP/Cbfb); Tg(Ly6aGFP/Cbfb) 14.5 dpc fetuses.B.Viability table; “hemorrhage” under the notes column refers to Cbfb-/-; Tg(Ly6a-GFP/Cbfb) conceptuses.
Figure S2 (related to Figure 5). Expression of Tg(Ly6a-GFP/Cbfb) in the 24s yolk sac.Scale bars 100 m. Figure represents the MAX of 30 consecutive 5 m z-sections for CD31and GFP.
Figure S3 (related to Figure 6). Expression of Tg(Tek-GFP/Cbfb) and Tg(Ly6a-GFP/Cbfb) inthe A U V of 11.5 dpc Cbfb / and Cbfb /- conceptuses.A.Scatter plot from the A U V of an 11.5 dpc conceptus indicating the gating strategy foranalyzing CD144 CD45- endothelial cells.B.Histograms illustrating the percentage of GFP/CBF endothelial cells in the A U V ofconceptuses carrying the individual transgenes, or both transgenes.C.Summary of data from individual conceptuses carrying transgenes. Error bars indicateSEM. The percentage of endothelial cells in A U V carrying both transgenes (Tek Ly6a) wassignificantly different from those expressing either transgene alone determined by one-wayANOVA.
Figure S4Figure S4 (related to Figure 7). Expansion of Tg(Ly6a-GFP) expression at 14.5 dpc.White bars indicate 50 m.A,B.Transverse section at level of the fetal liver. Red, CD34; Green, GFP; Ao, dorsal aorta,Nc, notochord; Es, esophagus.C.Ly6a-GFP expression in endothelial cells of the dorsal aorta.D.Fetal liver cells and vessel lining.E.Epithelial cells of the metanephric kidney.F.Lung macrophages.
Figure S5Figure S5. Ly6a expression is not dependent on endogenous CBF .A.Whole mount view of the umbilical vasculature and placenta of 11.5 dpc Cbfb /- andCbfb-/- conceptuses carrying the Ly6a-GFP/Cbfb transgene.B.Similar numbers of GFP/CBF cells in the Sca-1 population from the A U V. of 11.5dpc conceptuses.C.GFP/CBF cells in the A U V of 11.5 dpc Cbfb / and Cbfb-/- conceptuses carrying theLy6a-GFP/Cbfb transgene. Note that the percentage of GFP/CBF endothelial cells (CD45-
CD144 ) is equivalent in Cbfb / and Cbfb-/- conceptuses. The CD45 CD144- cells are primitivemacrophages which are largely GFP/CBF -, and are absent in Cbfb-/-; Tg(Ly6a-GFP/Cbfb)conceptuses.D.Average number of GFP/CBF cells in the A U V of 11.5 dpc Cbfb / or Cbfb /-conceptuses carrying the Ly6a-GFP/Cbfb transgene ( / or /-; Ly6a) versus Cbfb-/-;Tg(Ly6aGFP/Cbfb) conceptuses (-/-; Ly6a).E.Confocal image of GFP/CBF cells (arrow) in the endothelium of the dorsal aorta (Ao)of a 10.5 dpc Cbfb-/-; Tg(Ly6a-GFP/Cbfb) conceptus.
Cell Stem Cell, volume 9 Supplemental Information Erythroid/Myeloid Progenitors and Hematopoietic Stem Cells Originate from Distinct Populations of Endothelial Cells Michael J. Chen, Yan Li, Maria Elena De Obaldia, Qi Yang, Amanda D. Yzaguirre, Tomoko Yamada-Inagawa, Chris S. Vink, Avinash Bhandoola, Elaine Dzierzak, and Nancy A. Speck
