Epigenetic Control of MAGE Gene Expression by the KIT Tyrosine Kinase

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Epigenetic Control of MAGE Gene Expression by the KIT Tyrosine Kinase Bing Yang, Jianqiang Wu, Nityanand Maddodi, Yongsheng Ma, Vijayasaradhi Setaluri, B. Jack Longley Journal of Investigative Dermatology Volume 127, Issue 9, Pages 2123-2128 (September 2007) DOI: 10.1038/sj.jid.5700836 Copyright © 2007 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 1 Imatinib inhibits KIT-phosphorylation in HMC-1.1 cell line but not the HMC-1.2 cell line. Immunoprecipitation with anti-KIT and immunoblotting with anti-phosphotyrosine antibody (anti-PY) shows the regulatory type mutant-activated KIT in HMC1.1 is inhibited by Imatinib but the enzymatic site type mutant KIT expressed in the HMC1.2 cell line is resistant to Imatinib. Journal of Investigative Dermatology 2007 127, 2123-2128DOI: (10.1038/sj.jid.5700836) Copyright © 2007 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 2 Imatinib downregulates MAGE protein expression in sensitive HMC1.1 cell line but not in HMC1.2 which expresses Imatinib resistance. Immunoblotting shows MAGE-A and MAGE-C proteins are downregulated in HMC1.1 cells at 12 and 24hours after induction of apoptosis by Imatinib, but not downregulated in the Imatinib-resistant HMC1.2 cell line. Note that no significant expression of MAGE-B protein is identified. Note also that the HMC1.2 cell line does not express detectable MAGE-C2 protein, in agreement with the very low levels of MAGE-C2 mRNA seen in this cell line. Journal of Investigative Dermatology 2007 127, 2123-2128DOI: (10.1038/sj.jid.5700836) Copyright © 2007 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 3 KIT inhibition affects methylation of MAGE-A3 and MAGE-C2 promoters in Imatinib sensitive human mast cells. Methylation-specific PCR amplification of MAGE promoter regions from DNA extracted before or after addition of Imatinib, which inhibits activated KIT in the HMC1.1 mast cell line but not the HMC1.2 subclone. Each band confirms the presence of target DNA that is either unmethylated (U) or methylated (M). Journal of Investigative Dermatology 2007 127, 2123-2128DOI: (10.1038/sj.jid.5700836) Copyright © 2007 The Society for Investigative Dermatology, Inc Terms and Conditions