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. 2015 Sep 24;5(4):490–498. doi: 10.1016/j.stemcr.2015.08.014

Figure 3.

Figure 3

OCT4 Knockdown Eliminates Endoderm Differentiation

HES cells were transfected with siRNA against OCT4 or a control scramble siRNA construct 18 hr prior to initiation of endoderm differentiation as described in Figure 1.

(A) Flow cytometry analysis of OCT4 protein levels at D0, and T and EOMES levels at D1 of endoderm differentiation.

(B) QRT-PCR gene expression analysis of T and EOMES at D1 and FOXA1 and SOX17 at D3 of differentiation. Data shown are an average three independent experiments. Statistical significance represented as ∗p < 0.05.

(C) Flow cytometry analysis of CXCR4, FOXA1, and SOX17 at D3 of endoderm differentiation.

(D–F) ChIP-qPCR analysis comparing D1 differentiation cultures treated with OCT4 or scramble siRNA on primitive streak genes (T and EOMES) and endodermal genes (FOXA2 and SOX17). (D) Anti-EZH2. (E) Anti-SUZ12. (F) Anti-H3K27me3. Sites used for ChIP-qPCR are indicated in kilobases (K) from the transcription start site. Data shown are an average from three independent experiments. Statistical significance represented as ∗p < 0.05.