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. Author manuscript; available in PMC: 2012 Feb 1.
Published in final edited form as: Nat Chem Biol. 2011 Jul 3;7(8):531–537. doi: 10.1038/nchembio.598

Figure 5.

Figure 5

LID domain regulates transcription factors. (a) The LID domain was fused to the C-termini of six transcription factors that were independently transduced into NIH3T3 cells. Cell populations were treated with either vehicle or 2 μM Shield-1, and cell lysates were immunoblotted with antibodies against the indicated proteins (Oct4, Klf4), anti-HA (Sox2, Myc, Lin28) or anti-Flag (Nanog). α-Tubulin serves as the loading control (LC). Lane A represents untransduced NIH3T3 cells. Lanes B and C are from cells that were treated with vehicle (−) or with 2 μM Shield-1 for 24 h (+). Full western blots are shown in Supplementary Fig. 10. (b) Nuclear reprogramming of MEFs derived from transgenic mice encoding an Oct4 promoter driving GFP. MEF(Oct4/GFP) cells (ref. 24) were transduced with Sox2, Myc, Klf4, and Oct4 (left) or with Sox2, Myc, Klf4, and Oct4-LID (right). Cells were treated with vehicle or with 2 μM Shield-1 on day 3 post-infection. Alkaline phosphatase (AP) staining was performed on day 14 post-infection. Insert scalebars represent 5 mm. (c) GFP positive cells were transferred on day 19 to a new feeder layer. APC conjugated SSEA-1 (Stage-Specific Embryonic Antigen-1) antibody staining was performed on day 24 post-infection. Insert scalebars represent 50 μm.