Figure 1.

Kdm2b is highly expressed in mESCs and is a direct target of Oct4 and Sox2. (a) Relative expression levels of Kdm2b LF in ESCs, embryonic germ cells (EGCs), neural progenitor cells (NPCs), MEFs, C2C12 myoblasts and pancreatic β cells (Min6) measured by RT–qPCR. Data are mean ± s.d., n = 3. The source data for statistics are provided in Supplementary Table S5. (b) Relative expression levels of Kdm2b LF at different days of embryoid body differentiation measured by RT–qPCR and normalized to Gapdh. The mRNA level of the mESCs (D0) is set as 1. Data are mean ± s.d., n = 3. The source data for statistics are provided in Supplementary Table S5. (c) Western blot analysis of Kdm2b LF and Oct4 protein levels at different days of embryoid body differentiation. Tubulin serves as a loading control. Full scans of blots are shown in Supplementary Fig. S8a. (d) Bottom, ChIP–qPCR analysis demonstrated that Oct4 and Sox2 occupy the Kdm2b gene promoter. Data are presented as percentage of input DNA. The Kdm2b gene locus and the location of the various amplicons are indicated in the diagram at the top of the panel. The TSS and the relevant exons are indicated. Data are mean ± s.d., n = 3. The source data for statistics are provided in Supplementary Table S5. (e) An analysis of published ChIP-seq data⁴⁵ showing that Oct4 and Sox2 occupancy is enriched at the promoter of the mouse Kdm2b gene. (f) Presence of a consensus Oct4-binding sequence (highlighted) at the promoter region of mammalian Kdm2b genes. (g) Representative images show the morphology (top panels) and alkaline phosphatase activity (bottom panels) of control and Oct4 knockdown mESC cells. Scale bars, 100 μm. (h) Relative expression levels of Oct4 and Kdm2b in the control and Oct4 knockdown mESCs. The relative expression levels are normalized to Gapdh. The level of gene expression in the control knockdown mESCs is set as 1. Data are mean ± s.d., n = 3. The source data for statistics are provided in Supplementary Table S5.