Fig. 4. JMJD3 cooperates with KLF4 to activate target genes.

a Immunoprecipitation with anti-FLAG beads in nuclear extracts from OSKM-reprogrammed P2 MEFs with FLAG-tagged JMJD3 or Empty at day 5, followed by western blotting with the indicated antibodies. b Number of GFP⁺ colonies at day 14 in P2 MEFs reprogrammed with OSM (OCT4, SOX2, and c-MYC) and the indicated levels of KLF4 (high and low levels represent normal and 1/5 volume of retrovirus, respectively) plus Empty or JMJD3. c ChIP-qPCR for H3K27me3 at TSS and enhancer (Enh) of Sall4 in P2 MEFs reprogrammed with OSM and low or high level of KLF4 plus Empty or JMJD3 at day 5. d Number of GFP⁺ colonies in P2 MEFs reprogrammed with OSK (OCT4, SOX2, and KLF4) at day 6 or OS (OCT4 and SOX2) at day 20 plus Empty or JMJD3 in iCD1 medium. e Western blotting for E-cadherin (E-cad) in P2 MEFs transduced with the indicated constructs at day 5. f ChIP-qPCR for H3K27me3 at TSS of Cdh1 in P2 MEFs transduced with the indicated constructs in medium with Vc at day 5. g Schematic depiction of the EpiSC-to-naïve PSC (Epi-to-naïve) transition using KLF4 in OG2 EpiSCs (upper panel); images and number of GFP⁺ colonies and also percentage of GFP⁺ cells measured by flow cytometry at day 4 with Empty or JMJD3 (lower panel). Scale bar, 250 μm. h Schematic depiction of the Epi-to-naïve transition in Rex1GFPd2 epiblast-like cells (EpiLCs) using KLF4 (upper panel), the percentage of GFP⁺ cells measured by flow cytometry at day 4 with Empty or JMJD3 (lower panel). Error bars represent the s.e.m. Data are presented as mean ± s.e.m. from n = 3 b–d, f–h biologically independent experiments. Statistical analyses were performed using a two-tailed unpaired Student’s t-test (*P < 0.05; **P < 0.01; ***P < 0.001). P values: 2.72 × 10⁻⁵ b; 0.0002, 0.0432 c; 0.009 d; 0.0136, 0.0169 f; 0.0021, 0.0012 g; 0.0056 h. Experiments were repeated independently three times a or twice e with similar results. Source data are provided as a Source Data file.