Figure S2.

In vitro expansion and differentiation of the skin Ebf2⁺ and Ebf2⁻ PαS cell subsets. (A) Images of CFU-Fs derived from the Ebf2⁺ and Ebf2⁻ cell subsets. Scale bars are 500 µm. (B) PDT of the skin CD45⁻TER119⁻CD31⁻Ebf2⁺ and Ebf2⁻ stromal cells in culture. PDT was calculated based on culture time (CT)/cell doubling (CD) where CD = log (NH/NI)/log 2, NH is harvested cell number, and NI is the initial cell number. (C) Representative images of osteogenic, adipogenic, and chondrogenic differentiation of the skin Ebf2⁺ and Ebf2⁻PαS cells. Scale bars are 500 µm (osteogenic), 100 µm (adipogenic), and 20 µm (chondrogenic). (D) Bodipy 500/510 and oil red O staining of differentiated adipocytes from single skin Ebf2⁺ and Ebf2⁻PαS cells–derived CFU-F clones 21 d after the induction. Green and red represent adipocytes stained with bodipy 500/510 and oil red O, respectively. Scale bars are 100 µm. (E) Representative images of toluidine blue staining on micromass pellet after chondrogenic induction in vitro. The chondrogenic differentiation in a micromass pellet culture was performed on culture expanded BM MSCs and skin Ebf2⁺ and Ebf2⁻ cell subsets. Scale bars are 20 µm. Related to Fig. 2.