Expression of JAM-C on HSCs. (A) BM cells from BL6 mice were stained with anti-lineage cocktail (anti-CD3ϵ, anti-B220, anti-CD11b, anti–Gr-1, and anti-Ter119) and the lineage-positive (Lin⁺) and lineage-negative (Lin⁻) populations were analyzed for their JAM-C expression. (B) JAM-C expression on HSCs in the Lin⁻ population defined by expression of Sca-1 and c-Kit (LSK). (C) BM cells were individually stained with lineage markers (anti-CD3ϵ, anti-B220, anti-CD11b, anti–Gr-1, and anti-Ter119) and analyzed for their expression of JAM-C. (D) Blood, thymus, and spleen cells were stained for JAM-C expression on LSK cells. (E) Staining of the different stem cell populations contained within the LSK gate with anti–JAM-C: LT-HSCs (Lin⁻, Sca-1⁺, c-Kit⁺, Thy1.1⁺, and Flt-3⁻), ST-HSCs (Lin⁻, Sca-1⁺, c-Kit⁺, Thy1.1⁺, and Flt-3⁺), and MPPs (Lin⁻, Sca-1⁺, c-Kit⁺, Thy1.1⁻, and Flt-3⁻). (F) Expression of JAM-C on lymphoid progenitors (CLPs: Lin⁻, Sca-1^int, c-Kit^int, and Il7Rα⁺), and myeloid progenitors (CMPs: Lin⁻, Sca-1⁻, Il7Rα⁻, c-Kit⁺, CD34⁺, and FcγRII/III⁻; GMPs: Lin⁻, Sca-1⁻, Il7Rα⁻, c-Kit⁺, CD34⁺, and FcγRII/III⁺; MEPs: Lin⁻, Sca-1⁻, Il7Rα⁻, c-Kit⁺, CD34⁻, FcγRII/III⁻). Shown are representative histograms with anti–JAM-C staining (black histogram line), rabbit serum IgG (filled gray histogram), or anti–JAM-C together with 10 times excess of blocking murine JAM-C–Fc (hatched histogram line). The numbers above the brackets indicate the percentage of cells expressing JAM-C.