Fig. 3: GmLDL2 is an H3K4 demethylase.

a The affinity-purified recombinant His-GmLDL2 from E.coli was incubated with calf thymus histone for H3K4me1/2 demethylase activity assay, and the histone methylation status was determined using methylation-specific antibodies. His-HsLSD1 was used as the positive control, and anti-H3 was used as a loading control. The experiment was repeated three times independently with similar results. b Quantification analysis of (a) by ImageJ. Data represent means ± SD (n = 3 biological replicates). The P value was generated from a two-sided Student’s t-test. c GmLDL2-GFP exhibits H3K4me1/2 demethylase activity in N.benthamiana leaves. GmLDL2-GFP expression in N.benthamiana nuclei (arrows) was visualized by monitoring GFP fluorescence (green). The histone methylation status was analyzed by immunostaining with specific histone methylation antibodies (red). The location of nuclei was visualized using 4’,6-diamidine-2-phenylindole staining (blue). Scale bars, 50 μm. d Statistical analysis of the levels of fluorescent signals in (c) and Supplementary Fig. 5. Data represent means ± SD (n = 3 biological replicates). The P value was generated from a two-sided Student’s t-test. At least 200 nuclei expressing GFP, HsLSD1-GFP, or GmLDL2-GFP were analyzed in each biological replicate. Source data are provided as a Source Data file.