Fig. 2: The adult TCF21^lin cells are multipotent and can be directed to differentiate to Leydig and smooth muscle cells in vitro.

Schematic representation of experimental timelines for myoid (A) and Leydig cell (B) directed differentiation. Gating strategy used for FACS is presented in Supplementary Fig. 2G. C Immunofluorescence staining of smooth muscle actin (SMA) in clonal TCF21^lin cells after 7 days of in vitro culture in the presence of differentiation media. Representative images of n = 13 technical replicates of TCF21^lin/cKit⁻, n = 6 technical replicates of TCF21^lin/SCA1⁺/cKit⁻, n = 6 technical replicates of TCF21^lin/SCA1⁺/cKit⁻/CD105⁺, and n = 14 technical replicates of TCF21^lin/SCA1⁺/cKit⁻/CD105. Scale bar: 100 μm for all images in the panel. D Immunofluorescence staining of steroidogenic factor 1 (SF1) in clonal TCF21^lin cells after 14 days of in vitro culture in the presence of differentiation media. Representative images of n = 19 technical replicates of TCF21^lin/cKit⁻, n = 17 technical replicates of TCF21^lin/SCA1⁺/cKit⁻, n = 12 technical replicates of TCF21^lin/SCA1⁺/cKit⁻/CD105⁺, and n = 22 technical replicates of TCF21^lin/SCA1⁺/cKit⁻/CD105⁻. Scale bar: 100 μm for all images in the panel.