Skip to main content
. 2012 Sep 17;109(40):16202–16207. doi: 10.1073/pnas.1208533109

Fig. 2.

Fig. 2.

Zfp281 is required for Nanog autorepression and the integrity of the NuRD repressor complex in ESCs. (A) The strategy for inducible knockdown of Zfp281 in NG4 cells. (B) Flow cytometry analyses of Nanog-GFP reporter activity upon shRNA-mediated knockdown of Zfp281. Nanog-GFP was analyzed 3d after puromycin selection and Dox treatment. (C) RT-qPCR analyses of EndoNanog and Zfp281 expression in the samples described in B. (D) The strategy for inducible FLbioNanog expression in both Zfp281+/+ and Zfp281−/− ESCs. (E and F) RT-qPCR analyses of FLbioNanog and EndoNanog expression upon Dox treatment in Zfp281+/+ (E) and Zfp281−/− (F) ESCs. (G) Zfp281 is associated with the NuRD repressor complex in ESCs. Total peptide numbers identified by mass spectrometry are listed. (H) Confirmation of endogenous association of Zfp281 with Nanog and the NuRD proteins by immunoprecipitation (IP) and WB analyses in Zfp281+/+ and Zfp281−/− ESCs. (I) Zfp281 is required for the integrity of the NuRD repressor complex. (J) Interaction between Mta1/2 and Hdac2 is not affected by Zfp281 depletion.