Figure 1.

Loss of Dpf2 Affects ESC Self-Renewal and Leads to Increased Apoptosis and Cell-Cycle Defects

(A) Quantification of a colony-formation assay for WT, Dpf2^fl/fl, and Dpf2^−/− mouse ESCs. Given is the mean of three replicates and the SD. ^∗∗∗p < 0.001.

(B) Representative fluorescence-activated cell sorting (FACS) plots of Annexin V and 7-aminoactinomycin D (7-AAD) levels in Dpf2^fl/fl and WT control ESCs. Percentages of cells with different apoptosis marker levels are indicated in brackets.

(C) Alkaline phosphatase (AP) staining assay for Dpf2^fl/fl and Dpf2^−/− ESCs. Colonies were scored as undifferentiated (undiff), mixed, and differentiated (diff). The mean and SD of three replicates is displayed. ^∗p < 0.05, ^∗∗p < 0.01.

(D) Transcript levels of pluripotency-associated genes in Dpf2^fl/fl and Dpf2^−/− ESCs based on qPCR.

(E) Western blot for OCT4, SOX2, NANOG, and TBX3 protein levels in Dpf2^fl/fl and Dpf2^−/− ESCs; α-TUBB served as a loading control.

(F) Schematic of the affinity purification of FLAG-tagged DPF2 in ESCs and the MS procedure.

(G) DPF2-interacting proteins annotated in the STRING (Search Tool for the Retrieval of Interacting Genes/Proteins) database. Subunits of the BAF (blue), NuRD (tan), MCM (green), and MSH complexes (yellow) are highlighted.

(H) GO term and KEGG (Kyoto Encyclopedia of Genes and Genomes) pathway enrichment of DPF2-interacting proteins. Selected terms are shown. FDR, false discovery rate.