DEV02286 Supplementary Material

Files in this Data Supplement:

• Supplemental Table 1 (Adobe PDF) -

  Table S1. Primers used.

• Supplemental Figure 1 -

  Fig. S1. Morphology of NANOG overexpressing clones. Wild-type and NANOG overexpressing clones were seeded at a density of 2310⁴ cells per cm², either on feeders (upper panel) or in the absence of both feeders and CM (lower panel). The pictures were taken 4 days after seeding.

• Supplemental Figure 2 -

  Fig. S2. NANOG overexpressing cells adopt markers of primitive ectoderm cells. Real-time RT-PCR results described in Fig. 5B are shown here for each NANOG overexpressing clone separately. REX1: changes of 3.0- to -9.5-fold were observed (P<E–05). GBX2: the two clones that showed downregulation showed changes of five- to eightfold (P<0.0003). FGF5: fold changes of 3.5 to 15 were observed (P<0.02)

• Supplemental Figure 3 -

  Fig. S3. NANOG overexpressing clones show activation of the TGFb1/NODAL pathway. Genes whose expression levels changed in HESCs following treatment with MEDII were examined in the NANOG overexpressing clones. CRIPTO, LEFTYA and TBX1 were upregulated significantly (1.8 fold, P<0.0002; 6.75-fold, P<0.04; twofold, P<0.03, respectively), while FST was downregulated (2.45-fold, P<0.0001). However, no significant changes in the expression of GATA6 and ZNF1A1 were observed.