Extended Data Figure 3: The transcriptomes of D4 Nanog- and cytokine-induced PGCLCs are highly similar.
a, 100-200 ng/ml of Dox in EpiLCs results in NANOG expression levels similar to ESCs as shown by Western blot analysis for NANOG and α-TUBULIN (α-TUB) with GOF-GFP ESCs and D2 EpiLCs 24h after PGCLC induction (EpiLC aggregations) with Nanog (+Dox). For gel source data, see Supplementary Fig. 1.
b, PGCLC induction with 100 or 700 ng/ml Dox (for Nanog expression) with Noggin from GOF-GFP EpiLCs. Representative brightfield/GFP images at D4. GFP+ve cells are induced in both conditions. Scale bar, 200 μm.
c, Physiological (equivalent to ESCs) or higher levels of Nanog induce PGCLCs with comparable efficiency. FACS for GFP at D4 of PGCLC induction with 100 or 700 ng/ml Dox (for Nanog expression) with Noggin from GOF-GFP or Blimp1-GFP EpiLCs.
d, Alternative representation of qPCR data for Nanog, Prdm1 and Tfap2c shown in Fig. 1c. The induction of these genes in +cytokine conditions appears less evident, when compared to +Dox conditions. The data was log2-scaled, which allows a better comparison.
e, qPCR analysis of female GOF-GFP cells. GFP+ve cells were FACS-sorted. Note, the upregulation of PGC markers but not of the ESC marker Klf4. ΔΔCt mean values +/− s.d. (n=3 biological replicates). Colour code is shown in (d). Related to Fig. 1c.
f, qPCR analysis of male Blimp1-GFP cells. GFP+ve cells were FACS-sorted. Note, the upregulation of PGC markers but not of the ESC marker Klf4. ΔΔCt mean values +/− s.d. (n=3 biological replicates). Colour code is shown in (d). Related to Fig. 1c.
g, The transcriptomes of Nanog- and cytokine-induced PGCLCs are highly similar. Scatter plot showing the correlation of microarray data of ESCs, FACS-sorted D4 PGCLCs induced by cytokines or Nanog with Noggin. R indicates the Pearson correlation coefficient. n=2 biological replicates; related to Fig. 1d.
h, Nanog- and cytokine-induced PGCLCs cluster together as shown in unsupervised hierarchical clustering of microarray data described in (g). Related to Fig. 1d.
i, Heatmap showing the expression levels of selected genes from microarray data described in (g). Related to Fig. 1d.
j, Nanog-induced D4 PGCLCs are closely related to cytokine-induced D6 PGCLCs. PCA analysis with published microarray datasets5 (cross-platform comparison; see Methods for details). Note, that the separation of ESC samples is probably due to differences in genomic background and culture conditions.