Skip to main content
. Author manuscript; available in PMC: 2019 Apr 26.
Published in final edited form as: Nature. 2018 Oct 3;562(7728):595–599. doi: 10.1038/s41586-018-0581-5

Figure 1. Otx2 expression is down-regulated prior to expression of PGC TFs.

Figure 1

a. Scheme for PGCLC differentiation.

b. Top, scheme illustrating the time-points (hours) during PGCLC differentiation when mRNAs were analysed. Bottom, Q-RT-PCR of Otx2 and PGC TFs in E14Tg2a ESCs. Expression levels are normalised to TBP; h, hours; Values are means±SD, n= 3 biologically independent replicates.

c. Single cell quantification of immunofluorescence for Otx2 and Ap2γ in cytospin preparations of EpiLCs and cell aggregates at day 1 and day 2 of PGCLC induction. 2 biologically independent replicates were performed.

d. Whole mount immunofluorescence of E14Tg2a aggregates after 1 day of PGCLC differentiation. n=3. Scale bar, 50μm (top) and 10μm (bottom)

e-g. Representative confocal images of whole mount staining of embryos at pre-streak (e, n=4), early streak (f, n=3) and late streak (g, n=3) stages. Bar = 40μm (e), 100μm (f, g).

h-i. Magnified image of the regions highlighted in (f) and (g) respectively. OTX2-negative cells expressing BLIMP1 and FRAGILIS are outlined (g, h). Bar = 20μm.