a. Scheme for PGCLC differentiation, initiated from day 4 EpiLCs obtained after 1 passage from EpiLCs.
b. Representative morphologies and Oct4ΔPE::GFP expression from aggregates at day 6 of PGCLC differentiation from EpiLCs day4 (n=3 for 1 clone of each genotype). Bar = 200μm.
c. Scheme for generating chimaeras of GFP labelled Otx2+/+ or Otx2-/- ESCs with wildtype host embryos.
d. Comparison of the percentage contribution of GFP labelled wildtype (n=6) or Otx2-null ESCs (genotypes indicated, n=9 for each) to the PGC population in E7.5 chimaeric embryos. Each dot represents the percentage from one chimaera, centre lines and error bars represents means±SD. P value (two-sided unpaired T-test, 0.95 confidence intervals) is indicated. GFP positive cells were counted within the PGC population marked with BLIMP1 or SOX2 in each embryo.
e. Comparison of PGCs number in wild-type (n= 4) and Otx2-/- (n= 3) E7.5 embryos. PGCs were identified with Blimp1, AP2γ and Fragilis. Values= means ± SD.
f. Model indicating the point of operation of OTX2 during germline-soma segregation.
g. A scheme illustrating the regulatory relationships upstream and downstream of Otx2 during germline segregation.