(A) Schematic of embryonic day (E)14.5 skin with early stage hair follicle.
(B) Outline of cell sorting strategy from E14.5 Sox2GFP/Lef1-RFP double-transgenic skin. Diagram illustrates the back skin area microdissected for analysis.
(C) Immunofluorescence staining for E-cadherin (ECAD) marks all epithelial cells (Epi, orange arrow) and P-cadherin (PCAD) is highest in placode cells (Pc, red arrow). Dotted line demarcates basement membrane. DAPI highlights all nuclei.
(D) Sox2GFP/Lef1-RFP E14.5 back skin includes GFP+/RFP+ dermal condensate cells (DC, green arrow), GFP−RFP+ dermal fibroblasts (Fb, blue arrow), GFPlowRFP− Schwann cells (Sch, purple arrow); immunofluorescence for DCT confirms RFP+ cells in the epidermis are melanocytes (Mc, yellow arrow).
(E) FACS plots and gates for cell sorting. Starting from live cells, 8 distinct gates mark HF progenitors, niche cells, 4 other specific cell types and 2 mixed cell populations inclusive of the entire embryonic back skin.
(F) qRT-PCR for known marker genes demonstrates high enrichment for each purely isolated cell type. Data are mean ± SD from 2 measurements.
Scale bars are 50 μm in C, D. See also Figure S1.