Fig. 1: FACS isolation of eGFP⁺ cells.

a Schematic of the generation of the LMX1A-eGFP hESC reporter line. Biallelic insertion of eGFP into exon 9 of the LMX1A locus was performed using a nonstop mutation (TGA > TGG) in H9 hESCs and acted as a reporter for endogenous LMX1A protein expression. b Schematic overview of isolation and transcriptome analysis of eGFP⁺ cells on D20. c The eGFP⁺ fraction was determined according to fluorescence intensity in the eGFP channel using a 488-nm laser for excitation. d Bright-field images of unsorted, eGFP⁺, and eGFP⁻ cells at 6 h after cells were replated. e eGFP⁺ cells (green) also expressed LMX1A (red). f Flow cytometry analysis of EN1 and FOXA2 expression in eGFP⁺ cells on D22, with three biological replicates. Scale bars, 25 μm.