Fig. 1

NR2F2 expression during early human gonadal development and in COV434 and NT2/D1 cell lines. A The expression pattern of NR2F2 projected on the UMAP plot showing cell lineages in the scRNA-seq datasets of male and female somatic cells obtained from human gonads [24]. The color scale represents NR2F2 gene expression levels. Arrows indicate the predicted developmental trajectory of cell lineages based on evidence from the literature. B The monolayer differentiation protocol for human embryonic bipotential gonad described by Knarston et al. [25]. C, D RT-qPCR data of gene expression of NR2F2 transcript variants in CRL1502 (female) and PCS_201 (male) iPSCs (C), and of the bipotential gonad markers GATA4, ZFPM2, EMX2, and WT1, and the steroidogenic cell marker STAR (D) during the differentiation protocol. NR2F2 v3 was not detected under these experimental conditions. Data were normalized as a percentage of GAPDH (reference gene) expression. Mean ± SEM (n = 3). E RT-qPCR assay comparing the expression of NR2F2 transcript variants (v1–4) in COV434, NT2/D1, and HepG2 cell lines. Data is represented as a percentage of S8 (reference gene) expression. Mean ± SEM (n = 4). One-Way ANOVA followed by Tukey test (v1, v2, and v4) and t-test (v3). Asterisks indicate statistical significance in relation to COV434 expression. ***p < 0.001, ****p < 0.0001