Extended Data Fig. 3: EGLN2 promotes H3P16oh in vitro and in vivo.
From: Histone H3 proline 16 hydroxylation regulates mammalian gene expression
a–c, MS spectrum for synthetic hydroxylated peptide(a), identified hydroxylated (b) or non-hydroxylated (c) H3 peptide at P16 derived from biotin-H3(12-20) of in vitro hydroxylation assay. d, MS intensity of H3P16oh derived from biotin-H3(12-20) peptide of in vitro hydroxylation assay. e, Retention time of HPLC/MS analysis and the MS signal of biotin-H3(12-20) peptide with P16oh derived from in vitro hydroxylation assay and the synthetic peptide. f, Intensity value of biotin-H3(12-20) peptide with P16oh derived from in vitro hydroxylation assay. n = 2; mean ± s.d.; unpaired two-tailed Student’s t-test; *P < 0.05, **P < 0.01, ***P < 0.001. g, Immunoblots for lysates from MDA-MB-231 cells transduced with CRISPR-V2 sgctrl and sg13. h, Intensity value of endogenous histone peptide (KSTGGKAPR) with P16oh derived from MDA-MB-231 cells transduced with CRISPR-V2 sgctrl and sg13. i, The relative intensity of the hydroxylated peptide compared to the non-hydroxylated peptide with or without EGLN2 knockout. j, Dot blot analysis with indicated biotinylated peptides diluted with different concentrations and detected with either anti-H3P16oh or anti-Biotin antibody. k, Immunoblots of lysates from 293 T cells with indicated treatment condition for 12 h.
