Analysis of CD11c⁺ B220⁺ cells in (wt + Tcf1^−/−) mixed BM chimeras.

Spleen and thymus cell suspension of chimeric mice were stained with anti-CD45.2–Cy5. LDF obtained from the thymus was 4-color stained with anti-CD45.2–Cy5, anti-CD11c–FITC, anti-B220–CyChrome, and anti-F4/80–biotin AvPE. Results shown here are representative of 2 independent experiments. (A) Histograms show the reconstitution with Notch-1^−/− BM precursors in spleen, 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 Tcf-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 Tcf1^+/+BM precursors (gray bar) or Tcf-1^−/− BM precursors (black bar) in the thymic LDF. Numbers were obtained by multiplying the percentage of wt BM-derived cells (CD45.2⁻, 84%) or Tcf1^−/− BM-derived cells (CD45.2⁺, 16%) by the percentage of DCs or PDCs indicated in the respective quadrants.