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Figure 5: Effect of ATF7 on the LSD1/H3K9me2 signaling pathway. (a) MCF-10A cells were transfected with HER2-recombinant plasmid alone or with the combination of HER2- and ATF7-recombinant plasmids. Protein expression of HER2, ATF7, LSD1, H3K9me2, and H3K4me2 was determined by WB analysis. Relative quantitative analysis of ATF7 protein expression is shown on the right, (b) LSD1 activity as measured by the LSD1 Activity Quantification Assay Kits according to the manufacturer's instructions, (c) differential expression of LSD1 and H3K9me2 between HER2-negative and -positive breast cancer samples as determined by IHC staining. Scale bar, 50 μm, (d) gene expression profiles of ATF7 and KDM1A were obtained from the TCGA database. The association between ATF7 and ERBB2 in HER2-positive samples was determined by SPSS software (n = 79). *P < 0.05, **P < 0.01; ns = No significance. ATF7 = Activating transcription factor 7, HER2 = Human epidermal growth factor receptor 2, LSD1 = Histone lysine demethylase 1, H3K9me2 = Histone H3K9 dimethylation, WB = Western blot, IHC = Immunohistochemistry

Figure 5: Effect of <i>ATF7</i> on the LSD1/H3K9me2 signaling pathway. (a) MCF-10A cells were transfected with HER2-recombinant plasmid alone or with the combination of HER2- and <i>ATF7</i>-recombinant plasmids. Protein expression of HER2, <i>ATF7</i>, LSD1, H3K9me2, and H3K4me2 was determined by WB analysis. Relative quantitative analysis of <i>ATF7</i> protein expression is shown on the right, (b) LSD1 activity as measured by the LSD1 Activity Quantification Assay Kits according to the manufacturer's instructions, (c) differential expression of LSD1 and H3K9me2 between HER2-negative and -positive breast cancer samples as determined by IHC staining. Scale bar, 50 μm, (d) gene expression profiles of <i>ATF7</i> and <i>KDM1A</i> were obtained from the TCGA database. The association between <i>ATF7</i> and ERBB2 in HER2-positive samples was determined by SPSS software (<i>n</i> = 79). *<i>P</i> < 0.05, **<i>P</i> < 0.01; ns = No significance.<i> ATF7 </i>= Activating transcription factor 7, HER2 = Human epidermal growth factor receptor 2, LSD1 = Histone lysine demethylase 1, H3K9me2 = Histone H3K9 dimethylation, WB = Western blot, IHC = Immunohistochemistry