Fig 2. Epigenetically-defined hESC biomarkers have a role in pluripotency.

(A) Transcriptional activators (Expressed hESC-UnMe-GA-CGIs) and transcriptional repressors (Expressed hESC-Me-GA-CGIs) identified as functionally overrepresented hESC biomarkers. (B, C) Functional testing of transcriptional regulators in RH1 hESCs by siRNA knockdown. (B) RT-qPCR data showing log₁₀ fold change in expression of the siRNA-targeted gene, and associated effects on OCT4, NANOG and SOX2. Changes are relative to GAPDH expression, normalised to RH1 hESCs treated with negative control siRNA IDS-NULL. Asterisks indicate levels of statistical significance (unpaired t-test; *≤0.05, **≤0.01, ***≤0.001, ****≤0.0001). ND: Not Detected, even at 40 cycles of PCR. (C) Immunohistochemistry for NANOG and OCT4 72 hours after siRNA treatment. Scale bar = 100 μm.