Cd207

Cultivation periods and treatments were performed as described above (cultivation of epidermal explants with an α-RTN1A ab) in 96-well round bottom plates. Epidermal explants (6 mm in diameter/1 explant per well in triplicates per condition and time point) were removed and cells in the culture medium were collected from wells (Figure 3A and B). Cells were washed with PBS (Gibco, Thermo Fisher Scientific), stained with fixable viability dye and an ab cocktail for LC surface markers (FITC-conjugated CD207 [Beckman Coulter], CD1a [BD Pharmingen]), subsequently fixed, permeabilized, and stained with a secondary α-mouse cross-absorbed F(ab’) ab fragment conjugated with AF546 (Thermo Fisher Scientific) for the detection of primary abs. Samples were acquired using FACS Verse (BD Biosciences) and BD Suite software (v1.0.5.3841, BD Biosciences). Viable CD207⁺CD1a⁺ eLCs were enumerated (Figure 3A) and the signal of the primary ab measured and shown as MFI of AF546 (Thermo Fisher Scientific) (Figure 3B).