Figure S1. SOX17, TFAP2C, and BLIMP1 do not create human primordial germ-cell-like cells.

(A) The transgene expression levels in the designated hiPSC clones stimulated with (+) or without (−) Dox (1.0 μg/ml) for 24 h. Two clones were examined for each transgene combination. To quantify the transgene expression levels by qRT-PCR, primer pairs for the rabbit β-globin polyA sequence (rBGpA) or for the transgene coding sequences were used, and the ΔCt was calculated from the average Ct value of two housekeeping genes, RPLP0 and PPIA (set as 0). The expression levels of SOX17, TFAP2C and BLIMP1 in d2 human primordial germ-cell–like cells (BT⁺AG⁺ cells) measured by primer pairs for detecting their 3′ untranslated regions are shown as references. n.d., not determined. (B) Western blot analysis of the expression of SOX17, BLIMP1, and TFAP2C in the SOX17/TFAP2C/BLIMP1 hiPSC clone without (−) or with (+) 1.0 μg/ml of Dox for 24 h. α-TUBULIN was used as loading control. (C, D) Expression dynamics of rBGpA (transgenes) and the indicated endogenous genes in the TFAP2C/BLIMP1 (C) and SOX17/BLIMP1 (D) clones induced by bone morphogenetic protein 4 (black) or Dox (red). d1: whole aggregates; d2/d4: BT⁺AG⁺ cells for induction by bone morphogenetic protein 4, BT⁺ cells for induction by Dox. For each gene, the ΔCt was calculated from the average Ct value of two housekeeping genes, RPLP0 and PPIA (set as 0). The bars indicate the mean value of each time point. (C, D) Two independent experiments for the TFAP2C/BLIMP1 (C) and SOX17/BLIMP1 (D) clones were performed.

Source data are available for this figure.