Fig. 4

Identification of morphological changes with biomarkers, external features, and FCM. The cell portion of FasL⁻/AMH⁺ and FasL⁺/AMH⁺ cells in group mES+Trans were detected via FCM. The result of the whole cell population of mES+Trans between 13 and 48 days is shown in a. The result of the cells removed of PCs between 13 and 34 days is shown in b. Tests were performed every 7 days and expressed as mean ± SD (n = 3 independent experiments). c Identification of the transcriptional level of epithelial markers and mesenchymal markers reveals the timing of morphological transformation (MET or EMT) between 5 and 45 days. qPCR was performed every 5 days and expressed relative to the highest expression among each marker. qPCR results took the mean value and are shown in the heat map (n = 3 independent experiments). MET mesenchymal to epithelial transformation. EMT epithelial to mesenchymal transformation. Heat map indicates the gene expression levels of major transcription factors in group mES+Trans for d pluripotency, onset of gonads, bi-potential gonads, and male gonads, and for e introduced factors. qPCR results show changes in gene expression relative to the highest expression in each marker. Results were expressed as mean and transformed into heat map (n > 2)