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DNA 6mA Demethylase ALKBH1 Orchestrates Fatty Acid Metabolism and Suppresses Diet-Induced Hepatic Steatosis 1 Results 2 DNA 6mA Level Is Increased in Liver of Obese Mice 2 Expression of DNA 6mA Demethylase ALKBH1 Decreases in Mouse and Human Fatty Liver 3 Hepatic ALKBH1 Deficiency Aggravates Diet-Induced Hepatic Steatosis and Insulin Resistance 5 Transcriptomic and Chromatin Immunoprecipitation Analysis Suggest That ALKBH1 Regulates Lipid Metabolism 5 ALKBH1 Directly Binds to and Demethylates DNA 6mA Modification of Genes Involved in Lipid Uptake and Synthesis 6 Overexpression of ALKBH1 in Liver Protecting Against HFD-Induced Hepatic Steatosis and Insulin Resistance 6 Discussion 8 Materials and Methods 13 Animals 13 Primary Hepatocytes and Nonhepatocyte Isolation and Treatment 13 Nuclear and Cytoplasmic Protein Extraction 13 Gene Knockdown and Overexpression 13 Dot Blotting 13 Histology Analysis and Immunohistochemistry Staining 15 Glucose Tolerance Tests and Insulin Tolerance Tests 16 Quantification of Total TGs, FFAs, ALT, and AST in the Liver and Serum 16 RNA Isolation and Real-Time qPCR 16 Western Blot 16 RNA-seq Analysis 17 ChIP-seq Analysis 19 ChIP–qPCR Assay 19 Statistics 19

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UND

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nonfiction