Fig. 3.

Jumu is a 6mA-marked DNA-binding protein. a Logo representations of Forkhead domain (FHD)-binding motifs identified in the 6mA peaks at the 0.75-h stage. For 6mA peaks in 0.75-h stage embryos, the number and percentage of peaks containing the motif and the E-value of motif occurrence were indicated. b Western blot assays using the anti-Jumu antibody against purified complexes. Complexes purified by the 6mA-containing DNA probes exhibited much higher Jumu signals than those purified by the control probes. c Electrophoretic Mobility Shift Assay (EMSA) showing the signals of the shifted probe bands in gradient dose of Jumu protein. The signals were progressively increased with gradient dose of Jumu protein present in the reactions. d Competitive EMSA in different titrations showing the different affinities of recombinant FHD of Jumu protein to dA-dsDNA and 6mA-dsDNA. e The competitive ratio of dA or 6mA competitor in different titrations by the quantitative intensity of bands boxed (d). The one-tailed Student’s t test was used to analyze statistical variance. Data expressed as means of 4 independent experiments. Error bars indicate mean ± s.d. ***P < 0.001. f Surface plasmon resonance (SPR) measurements of the interaction between recombinant FHD of Jumu protein and dA/6mA-dsDNA. Equilibrium and kinetic constants were calculated by a global fit to 1:1 Langmuir binding model. RU, resonance units. g Overlap of 6mA enrichment peaks and Jumu-binding peaks in 0.75-h embryos. h The average signal profile of 6mA and Jumu-binding in common 6mA/Jumu peaks in 0.75- and 6-h stage embryos. Source data are provided as a Source Data file. n.s. not significant