Extended Data Fig. 9: Flow cytometric analysis of surface marker expression in transplanted Prrx1⁺ cells.

(a) Representative μCT results showed that transplanted Prrx1⁺ (Tomato⁺ITGAV⁺CD130⁺) cells underwent ossification. The scaffold was marked in red while the ossified matrix was in grey. Scale bar: 250 μm. (b) Flow cytometric analysis of transplanted BM Tomato⁺ cells. Tomao⁺ cells were extracted from the scaffolds after 3 months and analysed for expression of CD130 and other markers. (c) Flow cytometric analysis of transplanted iWAT or dermal Prrx1⁺ cells. Tomato⁺ cells were extracted from the iWAT or reconstituted skin and analysed for expression of CD130 and other markers. (d) Skin reconstitution experiments showed that Tomato⁺ITGAV⁺CD130⁺ cells isolated from the transplants could differentiate into different dermal cell types when re-transplanted. Since retrieved Tomato⁺ cells were not sufficient, only a portion of Tomato⁺ITGAV⁺CD130⁺ cells were used. The reconstituted skin samples were sectioned and immunostained with various antibodies. Upper panel: diagram for skin reconstitution experiment. Scale bar: 50 μm. (e) Flow cytometric analysis of re-transplanted BM or iWAT Tomato⁺CD29⁺CD130⁺ cells or dermal Tomato⁺ITGAV⁺CD130⁺ cells. Tomato⁺ cells were extracted from the implants after 3 months (for BM) or one month (for iWAT and dermis) and analysed for expression of CD130 and other markers. Each experiment was repeated three times independently with similar results obtained.