Figure 6

Differentiation of CD45.1⁺Lin⁻ cells into myeloid and granulocytic lineages in the inflamed skin and BM of the dermatitis mice.

(a) Flow cytometric analysis of Lin⁻ or Lin⁺ cells purified for transplantation. (b) Flow cytometric analysis of CD45.1⁺ cells in the inflamed skin of dermatitis mice transplanted with CD45.1⁺Lin⁻ or Lin⁺ cells (1 × 10⁶) at various time points after transplantation. CD45.1⁺ cells isolated from the skin were analyzed for expression of CD11b, Ly6G, Ly6C, F4/80 and CD11c on the designated days post-transplantation. Representative data gated for CD45.1⁺ cells are shown. Percentage values of each population within the CD45.1⁺CD11b⁺ cells and absolute cell numbers are shown. (c) DNCB-treated, CD45.1⁺Lin⁻ cell-transplanted and FTY-720-treated mice were injected with 1 mg BrdU daily for 3 days. FTY-720 was added to drinking water continuously until day 4 starting from day 1 post-transplantation. Leukocytes were isolated from the skin and stained with anti CD45.1-eFluor 450, anti-CD11b APC, anti-Ly6G PE and anti-BrdU-FITC antibodies. Representative data gated for CD45.1⁺BrdU⁺ cells are shown (d) Flow cytometric analysis of CD45.1⁺ cells in the BM. Data were analyzed as described above; representative data gated for CD45.1⁺ cells are shown. Data (a-d) are representative of three independent experiments (n = 5 mice/group/experiment). Data (B,d) are presented as means ±SEM. P values were determined using two-tailed unpaired Student’s t-tests; ns, not significant, *P < 0.05, **P < 0.01, ***P < 0.001.