Figure 10. DDB1 interacts with VprBP and DCAF11 in ESC and are required for ESC self-renewal.
(a-c) Ingenuity generated network of protein interactions of CUL4-DDB1 complex in (a) ESC, (b) 293T, and (c) HL-60. pink; DDB1 associated proteins (d) (left) immunprecipitation in 293T of endogenous VprBP protein followed by blotting for DDB1. (right) Transfection of Flag-DDB1 in 293T followed by immunopercipitation. (e-g) Nanog-GFP ES cells were transfected with pools of siRNAs under conditions of self-renewal and analyzed by FACS 72 hrs post-transfcetion. (e) Nanog-GFP expression. (n=3) (pink= proteins identified in ESC mass spectrometry analysis; blue=DDB1) (f) Bright field images of ESC colonies. (g) Representative experiment of relative expression of Nanog, Oct4, and Meox1. (n=3) (h) colony-forming units of LSK transduced with shRNAs. (i) Representative experiment of relative expression of following shRNA silencing. (j) Quantitative PCR of DCAFs in ESC and hematopoietic populations. (Differentiated= ESC differentiated for 48 hrs with retinoic acid). *p<0.05. **p<0.01. ***p<0.001.