Figure 5.
Role of BLIMP1 in hPGCLC Specification
(A) Western blot analysis of BLIMP1 and SOX17 in TNAP-positive (TNAP+) cells sorted from wild-type (WT) and BLIMP1 knockout (BLIMP1 KO) day 4 embryoids after hPGCLC induction. TUBULIN was used as loading control.
(B) FACS analysis of TNAP and NANOS3-mCherry on WT and BLIMP1 knockout (BLIMP1 KO) day 4 embryoids.
(C) Immunofluorscence for OCT4 and SOX17 in cryosections of WT and BLIMP1 KO day 4 and 8 embryoids. OCT4-positive cells are highlighted. Scale bar, 50 μm.
(D) Expression analysis by RT-qPCR for WT TNAP/NANOS3-mCherry double-positive cells (WT; TNAP+N3+) and BLIMP1 KO TNAP single-positive cells (BLIMP1 KO; TNAP+) sorted from day 4 embryoids. Relative expression levels are shown with normalization to β−ACTIN. Error bars indicate mean ± SD from two independent biological replicates.