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MiR-197-3p reduces bortezomib resistance in multiple myeloma by inhibiting IL-6 expression in a MEAF6-dependent manner 1 1 Introduction 1 2 Materials and methods 2 2.1 Ethics statement 2 2.2 Cell culture 2 2.3 qRT-PCR for detecting RNA expression 2 2.4 MTT assay for analyzing cell viability and BTZ IC50 2 2.5 Cell transfection 2 2.6 Flow cytometry for analyzing cell apoptosis 2 2.7 Western blotting for detecting protein expression 2 2.8 Chromatin immunoprecipitation (ChIP) for verifying the binding of MEAF6 to IL-6 promoter 2 2.9 RNA-binding protein immunoprecipitation (RIP) for verifying the binding of miR-197-3p to MEAF6 mRNA 3 2.10 Dual-luciferase reporter assay for verifying the binding sites between miR-197-3p and MEAF6 mRNA 3 2.11 Xenograft mouse model for assessing the function of miR-197-3p in vivo 3 2.12 Statistical analysis 3 3 Results 3 3.1 Low expression of miR-197-3p in BTZ-resistant MM cells 3 3.2 MiR-197-3p reduces BTZ resistance of MM cells 4 3.3 MiR-197-3p decreases the expression of IL-6 by inhibiting histone H3 acetylation 4 3.4 MiR-197-3p inhibits histone H3 acetylation in IL-6 promoter by targeting MEAF6 5 3.5 IL-6 counteracts the inhibitory effect of miR-197-3

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Language

UND

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nonfiction