Figure 5. LC replenishment after UV induced inflammation depends on PU.1 but not ID2. (A) Schematic representation of the experimental plan. (B–E) Mice reconstituted with BM of the indicated genotype were exposed to UV and the repopulation of the epidermis by Ly5.2+ inflammatory LCs was assessed by flow cytometry after 3 wk. (B) Contour plot shows the expression of CD11c and MHCII in ear epidermal cells derived from wild-type (wt) donor (Ly5.2+) and host (Ly5.1+). Numbers represent the frequency of the gated populations. (C) Chart represents the mean frequency of chimerism ± SD for the indicated genotype in the ear, trunk, and LNs (n = 4/genotype). (D) Contour plot shows the expression of CD11c and MHCII in epidermal cells derived from donors of the indicated genotypes. Graphs show the mean frequency ± SD of donor-derived LCs found in the host epidermis from four mice per genotype. (E) Top left, expression of CD11c and Langerin in donor (Ly5.2+) cells of the indicated genotype from peripheral LNs. Bottom left, dot plots show the expression of CD103 and Langerin within Ly5.2+CD11c+MHCIIhigh cells. Numbers represent the frequency of the gated populations. Right, bar graphs show the mean proportion ± SD of either CD11c+Langerin+ LN cells (gated as in top left) or CD11c+Langerin+CD103− LCs (gated as in bottom left) derived from the indicated genotype. Data in B–E are representative of two independent experiments with at least four independent chimeric mice per genotype. **, P < 0.01; ***, P < 0.001 (unpaired Student’s t test) compared with the wild-type sample. (F and G) Ly5.1 mice were reconstituted with BM from Id2cKO and exposed to UV. The repopulation of the epidermis by Ly5.2+ inflammatory LCs was assessed by flow cytometry after 6 wk. Contour plots show the expression of CD11c and MHCII in cells derived from donor (Ly5.2+, Id2cKO) and host (wt, ly5.1+) in the trunk. Numbers represent the mean frequency of LC (n = 3 ± SD) for the indicated compartment. (G) DNA was extracted from wild-type (Ly5.1+) and Id2cKO (Ly5.2+) BM-derived LC, gated as in F. The ID2 deletion efficiency was assessed by PCR. ID2cKO, CD11cCre−, Id2fl/fl and a wild-type control are shown. H2O, no DNA added to the reaction. The position of the wild-type, loxP flanked (fl), and deleted (Δ) alleles are indicated. Numbers on the left indicate the molecular weight in base pairs. Data are representative of two independent experiments.
Figure 5.

LC replenishment after UV induced inflammation depends on PU.1 but not ID2. (A) Schematic representation of the experimental plan. (B–E) Mice reconstituted with BM of the indicated genotype were exposed to UV and the repopulation of the epidermis by Ly5.2+ inflammatory LCs was assessed by flow cytometry after 3 wk. (B) Contour plot shows the expression of CD11c and MHCII in ear epidermal cells derived from wild-type (wt) donor (Ly5.2+) and host (Ly5.1+). Numbers represent the frequency of the gated populations. (C) Chart represents the mean frequency of chimerism ± SD for the indicated genotype in the ear, trunk, and LNs (n = 4/genotype). (D) Contour plot shows the expression of CD11c and MHCII in epidermal cells derived from donors of the indicated genotypes. Graphs show the mean frequency ± SD of donor-derived LCs found in the host epidermis from four mice per genotype. (E) Top left, expression of CD11c and Langerin in donor (Ly5.2+) cells of the indicated genotype from peripheral LNs. Bottom left, dot plots show the expression of CD103 and Langerin within Ly5.2+CD11c+MHCIIhigh cells. Numbers represent the frequency of the gated populations. Right, bar graphs show the mean proportion ± SD of either CD11c+Langerin+ LN cells (gated as in top left) or CD11c+Langerin+CD103− LCs (gated as in bottom left) derived from the indicated genotype. Data in B–E are representative of two independent experiments with at least four independent chimeric mice per genotype. **, P < 0.01; ***, P < 0.001 (unpaired Student’s t test) compared with the wild-type sample. (F and G) Ly5.1 mice were reconstituted with BM from Id2cKO and exposed to UV. The repopulation of the epidermis by Ly5.2+ inflammatory LCs was assessed by flow cytometry after 6 wk. Contour plots show the expression of CD11c and MHCII in cells derived from donor (Ly5.2+, Id2cKO) and host (wt, ly5.1+) in the trunk. Numbers represent the mean frequency of LC (n = 3 ± SD) for the indicated compartment. (G) DNA was extracted from wild-type (Ly5.1+) and Id2cKO (Ly5.2+) BM-derived LC, gated as in F. The ID2 deletion efficiency was assessed by PCR. ID2cKO, CD11cCre−, Id2fl/fl and a wild-type control are shown. H2O, no DNA added to the reaction. The position of the wild-type, loxP flanked (fl), and deleted (Δ) alleles are indicated. Numbers on the left indicate the molecular weight in base pairs. Data are representative of two independent experiments.

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