Figure 4. Identifying cDC1 in collagenase digested and spontaneous migrating populations.

Cells were isolated by collagenase digestion or spontaneous migration and stained as descried in Fig. 3. After enzymatic digestion cDC1s could be identified by gating on either the CD141⁺CD14⁻ (A) or the CD141⁺CD11c⁻ population (B); both of which expressed CLEC9A but not MR. (C and D) Sorted cDC2s and CD14⁺ MDMs isolated via enzymatic digestion were cultured for 24 h then stained for CD141⁺ (C) and compared with levels from the freshly sorted populations and CD11c and CD1a (D). Gating of CD141 DCs based on CD14 expression (CD141⁺CD14⁻) was compared with gating based on CD11c expression (CD141⁺CD11c^low). (E) CADM1 expression on cDC1 (purple) compared with all other blood or dermal DC (gray). (F) CLEC9A and MR surface expression of cDC1 and cDC2 in migrated populations.