Analysis of CD11c⁺ B220⁺ cells in (wt + Notch-1^−/−) mixed BM chimeras.

PBLs and thymi of Notch-1^+/+ chimeric mice were pooled and stained with anti-CD45.2–Cy5. LDF was also obtained from a pool of thymi that was 4-color stained with anti-CD45.2–Cy5, anti-CD11c–FITC, anti-B200–CyChrome, and anti-F4/80–biotin AvPE. Results are representative of 2 independent experiments. (A) Histograms show the reconstitution with Notch-1^−/− BM precursors in PBLs, total thymus, and thymic LDF. Contour plots show CD11c versus B220 expression (after gating out F4/80⁺ cells), by thymic LDF derived from wt BM precursors (CD45.2⁻, R1) or from induced Notch-1^−/− BM precursors (CD45.2⁺, R2). Numbers indicate the percentage of cells in the respective quadrant. (B) Bar diagram shows the relative proportion of DCs (CD11c⁺ B220⁻) and PDCs (CD11c⁺B220⁺) derived from wt BM precursors (gray bar) or induced Notch-1^−/− BM precursors (black bar) in the thymic LDF. Numbers were obtained by multiplying the percentage of wt BM-derived cells (CD45.2⁻, 52%) or induced Notch-1^−/−BM-derived cells (CD45.2⁺, 48%) by the percentage of DCs or PDCs indicated in the respective quadrants.