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. Author manuscript; available in PMC: 2016 Dec 3.
Published in final edited form as: Cell Stem Cell. 2015 Oct 29;17(6):689–704. doi: 10.1016/j.stem.2015.09.005

Figure 1. ZFP217 is required to maintain the pluripotent state of ESCs.

Figure 1

(A) RT-qPCR analysis of Zfp217 expression in MEFs, iPSCs and ESCs. Data are represented as mean ± SD; n = 3. ***p<0.0005; **p<0.005 versus MEFs.

(B and C) RT-qPCR analysis of Zfp217, Pou5f1, Sox2, and Zfp617 during RA-induced differentiation (B) and EB formation (C). Data are represented as mean ± SD; n = 3.

(D–E) RT-qPCR analysis of Zfp217 (D) and representative western blot (E) for ZFP217 upon Nanog, Pou5f1 and Sox2 depletion. The expression of the pluripotency factors was determined to monitor the knockdown efficiency. Error bars show ± SD; n = 3. ***p<0.0001 versus control shRNA. For the immunoblots β-ACTIN was used as a loading control. * non specific.

(F) RT-qPCR (upper panel) and western blot analysis (lower panel) to monitor Zfp217 knockdown efficiency. Error bars show ± SD; n = 3. ***p<0.0001 versus control shRNA. For the immunoblot, β-ACTIN was used as a loading control.

(G–H) AP staining of control and Zfp217-depleted ESCs (G). Percentage of ESC colonies were counted and depicted in the graph (H). Undifferentiated (UD); partially differentiated (PD); fully differentiated (FD).

(I) Proliferation rate of control and Zfp217-depleted ESCs relative to shRNA control at day 8. Error bars show ± SD; n = 3. *p=0.01 versus control shRNA.

(J) Cell cycle distribution of control and Zfp217-depleted ESCs examined by DNA content index. Error bars show ± SD; n = 3. ns= not significant; *p<0.05 versus control.

(K) Percentage of early apoptotic cells in control and Zfp217-depleted ESCs defined as Annexin V-positive and 7AAD-negative cells. Error bars show ± SD; n = 3. **p<0.005; ***p<0.001 versus control shRNA.