Fig. 2

Development fate identification. a Stepwise development of progenitor cells to eSCs. b Heat map of stage-specific marker expression of different groups. In each transduction group, a different factor was removed from the pool of six factors. Group All factors was transduced with all the six factors. Group No factor was the control group (mES+MEF) without transfection. Group −Sry was the mES cells transduced with the six factors except for Sry. The rest were constructed in the same manner. Flow cytometry (FCM) with specific markers indicating different cell stages. AMH⁺/Emx2⁺ cells indicated coelomic epithelium. AMH⁺/SF1⁺ cells indicated SF1-positive precursor cells. AMH⁺/FasL⁺ cells indicated eSCs. Fluorescence antibody staining was performed after the samples had cell membrane perforation with Triton X-100. The results were expressed as mean value of the maximum positive cell portion of each test group in 35 days (n > 3 independent experiments). Gene expression levels of transcription factors c lin28, d FOG2, e dhh, and f Sox8 were detected by qPCR. Results show changes in gene expression relative to the highest expression in each group. c–f were expressed as mean ± SD (n = 3 independent experiments). Asterisks indicate statistical significance of differences in the mean of gene expression by one-way ANOVA according to the positive group (All factors) (*P value < 0.05, **P value < 0.01, ***P value < 0.001)