Supplementary Figure 3: Overexpression of PAX5 and PRDM1 enhances germ cell potential of hESCs during in vitro differentiation.

(a) RT-qPCR analysis of expression level of PAX5 in H1 ESCs, PAX5 KO and cells overexpressing PAX5. Data are represented as mean ± SD of three replicates. (b) Bright field view of cells PAX5 OE and PRDM1 OE cells. Scale bars represent 50 µm. Experiments were independently repeated a minimum of three times with similar results. (c) Immunostaining of OCT4, PAX5 and PRDM1 in PAX5 and PRDM1 overexpression hESCs. OE represents overexpression. Scale bars represent 25 µm. Immunostaining experiments were independently repeated a minimum of three times with similar results. (d) RT-qPCR analysis of control, PAX5 OE and PRDM1 OE H1 hESCs before and after BMPs-induced differentiation. Data are represented as mean ± SD of n=3 independent replicates. (e) Genome browser representation of ChIP-seq tracks for PAX5 at the SYCP3 and SYCP1 loci. ChIP-seq were independently repeated twice with similar results. (f) RT-qPCR analysis of control and PAX5&PRDM1 double OE H1 hESCs after BMPs-induced differentiation. Data are represented as mean ± SD of n=3 independent replicates. Source data for a, d and f are in Supplementary Table 2.