Figure 3.

PRDM1 Is Required for BMP4- and WNT3A-Induced Germ Cell Formation in hESCs

(A) Immunofluorescence staining of H9 hESCs treated with BMP4 (100 ng/mL) and WNT3A (50 ng/mL) for the indicated days using DAPI and the indicated antibodies. Scale bar, 30 μm. Arrows indicate PRDM1⁺/OCT4⁺ cells.

(B) qRT-PCR analysis showed the expression of the indicated mRNA in H9 hESCs exposed to BMP4 and WNT3A treatment for the indicated days. Relative fold was compared with spontaneously differentiated cells without BMP4 and WNT3A treatment.

(C) qRT-PCR analysis showed the expression of the indicated mRNA in H9 hESCs treated with BMP4 (B) and WNT3A (W) in the presence or absence of NOGGIN (250 ng/mL) and DKK1 (500 ng/mL) for 5 days. Relative fold was compared with the levels of mRNA indicated in cells without treatment with BMP4 and WNT3A (mock).

(D) qRT-PCR for the indicated mRNA isolated from BMP4- and WNT3A-treated shControl or shPRDM1-expressing H9 cells on day 10. Relative fold was compared with the shControl group.

(E) Immunofluorescence staining of DAPI, PRDM1, and VASA in control and PRDM1-KD (GFP⁺) cells 5 days and 10 days after BMP4 and WNT3A treatment. Scale bar, 30 μm. The values in (B)−(D) are represented as the mean ± SEM of three independent experiments.

See also Figure S2.