Supplementary Figure 4: Lineage potential of thymus at E11.5.
From: Initial seeding of the embryonic thymus by immune-restricted lympho-myeloid progenitors
Related to main Figures 4 and 5. (a) Morphology of representative myeloid cells from cultures of E11.5 embryonic thymus. White arrows indicate monocytic cells, and black arrows granulocytic cells. Asterisk indicates OP9/OP9-DL1 stromal cells. Scale bars: 10 µm. (b) Representative acetylcholinesterase stained CFU-Mk colonies from E11.5 fetal liver. Scale bars represent 100 µm. (c) Mean (s.d.) number of DAF+ erythroid colonies generated from E11.5 unfractionated fetal liver cells cultured in semi-solid methylcellulose (n = 4; 50,000 cells/biological replicate). (d,e) Representative flow cytometry analyses of T cell (d) and B cell (e) producing cultures of individual thymic lobes from Rag1-GFP E11.5 embryos. (f) Gating strategy for FACS sorting of Lin–CD45loVE-Cad+c-Kit+ AGM stem/progenitor cells (HSPC). Numbers represent mean frequencies of total cells, from 3 litters analysed in 2 independent experiments. (g) Representative FACS profiles of pooled E12.5 thymuses. Virtually all Rag1-GFP+ cells express CD45 and progenitor markers c-Kit and Flt3 but are negative for lineage markers and CD25. The number in the first plot reflects the percentage of total cells, whereas subsequent plots show frequencies relative to total GFP+ cells. All frequencies are means of 3 litters (6-7 embryos pooled in each litter) from 2 independent experiments.
