Fig 2: ChIP-qPCR profiles of NuRD proteins, CBP proteins, and Gata1/Fog1 to the a-globin promoter during MEL cell differentiation. (A) Schematic representation of the a-globin promoter with ChIP-qPCR probes (1–5). GATA; Gata1 binding site, [CNRG-N(5-6)-CNRG]; CP2c binding site, CCAAT; Nfy binding site, TATA; TATA box. ChIP-qPCR profiles of NuRD proteins (B), CBP proteins (D), and Gata1 and Fog1 (E) on the a-globin promoter in uninduced WT (d0 WT), uninduced Mbd2 KD (d0 Mbd2 KD), and HMBA-induced WT (d3 WT) MEL cells. n = 2. IgG and Errc3 are used as internal negative controls. (C) Immunoblot analysis of the Ac-Hdac1 and Hdac1 expression during HMBA-induced differentiation of Mbd2 KD and Mbd3 KD MEL cells. Values are normalized to ß-tubulin and relative values over WT at d0 are shown as mean ± SEM; n = 2.

Fig 3: Western blot analysis of FOXM1, MBD3, CCND1, KLK10 and CASP2 expression in the HCT116 cells.Protein expression levels of FOXM1, MBD3, CCND1, KLK10 and CASP2 in HCT116 cells transfected with either 30nM of miR-8073 mimic or microRNA negative control (miR-NC) detected by Western blots. The images were numerated by software and ratios of the protein expression signal of miR-8073 treated cells over miR-NC treated cells were calculated.

Fig 4: NuRD is required to maintain a stable pluripotency state in hiPSCs A) MDS plot made from RNA-seq data of wild type, MBD3-KO and Rescued hiPSCs across a neural differentiation timecourse. Each point represents a biological replicate, and shapes indicate the days of differentiation. B) Heat map of genes found to be differentially expressed between WT and MBD3-KO cells in self-renewing conditions (day 0; FDR 5%). C) GO terms associated with genes significantly activated (open bars) or repressed (filled bars) in MBD3-KO cells relative to WT hiPSCs in self-renewing conditions.

Fig 5: NuRD facilitates an appropriate transcriptional response in mEpiSCs. A) An outline of the experiment, as in Fig. 4. B) Phase contrast images of wild type or Mbd3-KO mEpiSCs in self-renewing conditions. C) MDS plot of gene expression data collected across the neural differentiation time course, as in Fig. 3A. D) Heat map of DEG (FDR 1%) separated into four clusters by K-means clustering. E) Mean expression and most significant GO terms for each cluster as in Fig. 4B. A full list of GO terms and pathways is available in Tables S6 and S7. Gene expression changes during neural differentiation of an independent pair of WT and Mbd3-KO EpiSCs, verifying the results of the RNA-seq shown here, is displayed in Fig. S4.