Extended Data Fig. 9: The effect of hypoxia on EGLN2-H3P16oh-KDM5A-H3K4me3 signaling axis depends largely on the inhibition of EGLN2 and H3P16oh.
From: Histone H3 proline 16 hydroxylation regulates mammalian gene expression
a, Scatter plot showing the correlation of replicated (R1/2) CUT&RUN signals. b, Immunoblots of H3P16oh and HIF1α level in MDA-MB-231 cells with hypoxia treatment (1% O2) with indicated time. The values listed below the blots indicate the relative H3P16oh and H3K4me3 protein levels with H3 normalization, the samples derive from the same experiment and that blots were processed in parallel. c, qRT-PCR quantification of top 10 genes from MBA-MD-231 cells with hypoxia (1% O2) treatment for 30 min. d, qRT-PCR quantification of H3P16oh top 10 genes from MBA-MD-231 cells with siEGLN2 and siEGLN2 plus hypoxia(1% O2) treatment for 30 min. e, ChIP-qPCR to examine H3K4me3 binding to the promoter regions of the indicated genes with siEGLN2 and siEGLN2 plus hypoxia (1% O2) treatment for 30 min. f, qRT-PCR quantification for top 10 genes from MBA-MD-231 cells with siEGLN2 and siEGLN2 plus hypoxia (1% O2) treatment for 8 h. g, ChIP-qPCR to examine H3K4me3 binding to the promoter regions of the indicated genes with siEGLN2 siEGLN2 plus hypoxia (1% O2) treatment for 8 h. For panel c-g, the y axis shows averaged signals after normalization to indicated control (n = 3; mean ± s.d.; unpaired two-tailed Student’s t-test). *P < 0.05, **P < 0.01, ***P < 0.001.
