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. 2021 Jan 8;40(7):1231–1241. doi: 10.1038/s41388-020-01590-8

Fig. 5. MMSET modifies the histone methylation on DKK1 gene promoter.

Fig. 5

a Luciferase assay of DKK1-luc reporter in MM cells electroporated with plasmids encoding CREB and MMSET full length (FL), depletion of SET domain (ΔSET), or tyrosine to alanine mutation at 1179 amino acid site governing MMSET methyltransferase activity (Y1179A). b The MMSET and histone H3K36me2 levels in MM cells under normoxia or hypoxia conditions. c The enrichment of CREB and MMSET on the promoter of DKK1 in MM cells in hypoxia condition by ChIP-qPCR, and GAPDH gene was used as negative control. **P < 0.001 by Student’s t-test for n = 3 independent experiments. Co-immunoprecipitation shows d interaction between MMSET and CREB in MM cells under hypoxia condition for 24 h, and (e) in HEK239T cells ectopically expressed MMSET-3flag and wild type CREB (WT) or a constitutively active CREB mutant at S133D (CA). f Levels of H3K36me2 on the promoter of DKK1 gene in MM cells under hypoxia for up to 24 h by ChIP-qPCR, and g shows H3K36me2 levels on the promoter of DKK1 gene in MM cells exposure to hypoxia in the presence or absence of LW6 (10 μM), SB203580 (0.5 μM), or KG-501 (10 μM) for 24 h. GAPDH gene was used as negative control for both experiments. **P < 0.001 by Student’s t-test for n = 3 independent experiments.