Figure 6.

BMEx overlay potentiates BMP4 signaling and increases expression of critical PGC specification factors

(A) Volcano plot showing DEGs between hPSCs at day 0 and 2-differentiated progenitors with BMEx overlay.

(B) Immunofluorescence for TFAP2A, SOX17, and PRDM1 at days 2, 3, and 5 with BMEx overlay in line M54. TFAP2A is shown on top as a single channel. Dashed box is magnified (below), showing separate channels. Scale bars: 50 μm.

(C) Immunofluorescence for pSMAD1/5/9, GATA3, and SOX17 at days 2, 3, and 5 with or without BMEx overlay in line M54. pSMAD1/5/9 is shown on top as a single channel. Scale bars: 50 μm.

(D) Immunofluorescence for TFAP2A, EOMES, and SOX2 in line F99 at day 2 without (a) or with (b) BMEx overlay or with addition of 10 ng/mL activin A (C) or 10 μM SB431542 (D). Scale bars: 50 μm.

(E) Violin plots depict the quantification of the images in (D) as the mean fluorescence intensity in arbitrary units (A.U.) of TFAP2A (left) and EOMES (middle) in DAPI segmented areas (normalized to 1) per cell at day 2. The correlation between these two values per cell per condition was visualized in a scatterplot (right).

(F) Cartoon summarizing the hPGCLC differentiation progression in the BMEx overlay method as well as the perturbations tested (from D and E), with key analyzed markers depicted.

See also Figure S4.