FIG. 2.

WNT and RA act synergistically to specify TBX18⁺/WT1⁺ cell fate. (A) qRT-PCR analysis of TBX18 and WT1 expression in D14 cultures following treatment with 5 μM CHIR and the indicated concentrations of RA. BMS493, 5 μM. Gene expression was normalized to TBP. Error bars represent SEM; n = 3; *p < 0.05, **p < 0.01 versus CHIR (5 μM)-treated cultures, analyzed by the Student's t-test. (B) Flow cytometry analysis of the proportion of WT1⁺ cells in D14 cultures. Error bars represent SEM; n = 3. (C) High-content imaging assays of the proportion of TBX18⁺/WT1⁺ cells in D14 cultures treated with CHIR, RA/CHIR, or BMS493/CHIR. Error bars represent SEM; n = 3; **p < 0.01 versus CHIR (5 μM)-treated cultures. (D) Immunofluorescence staining of TBX18 and WT1 in D14 cultures treated with CHIR, RA/CHIR, or BMS493/CHIR. Yellow in the inset panels indicates TBX18 and WT1 coexpression. Scale bars, 100 μm. Color images available online at www.liebertpub.com/scd