Fig. 4.

Defective LC migration in LC/EpCAM-cKO mice in vivo. (A) Apparent retention of EpCAM-deficient LC in epidermis after TNCB application. Flow cytometry of CD45⁺ MHC Class II⁺ Langerin⁺ cells in ear epidermal cell suspensions prepared 36 h after 1% TNCB painting revealed a subpopulation of MHC Class II^high CD86^high activated LC in situ (cells to the right of the vertical dashed line; thick line, KO; thin line, WT; gray line with shading, isotype control). (B) Bar graph demonstrating transient retention of activated LC in ear epidermis of LC/EpCAM-cKO (KO) mice 36 h after TNCB painting. Data depicted represent means ± SEM of two or three experiments. n = 3 mice per group. *P < 0.005. (C) Attenuated migration of EpCAM-deficient LC from epidermis to draining LN. Abdominal skin of KO and WT animals was painted with 1% TRITC in dibutyl phthalate:acetone (1:1) or with vehicle, and single-cell suspensions were prepared from skin-draining LN and were analyzed by flow cytometry. Dot plots reveal TRITC⁺ cells within live-gated CD11c⁺ cells after 96 h. (D) Reduced absolute numbers of TRITC⁺ cells within live-gated CD11c⁺ MHC Class II⁺ Langerin⁺ CD103⁻ CD11b⁺ cells (LC) in LC/EpCAM-cKO mice based on initial cell counts from pooled skin-draining LN at 72, 96, and 120 h after TRITC painting. Data depict means ± SEM from three experiments. n = 3 mice per group. *P < 0.001.