Irisin directly stimulates osteoclastogenesis and bone resorption in vitro and in vivo: RNAseq dataset
Data files
Oct 16, 2020 version files 1.46 MB
Abstract
Methods
Total RNA was isolated with Trizol reagent and RNeasy mini kit from primary bone marrow progenitors induced to osteoclast differentiation with RANKL and MCSF, cultured with or without 10 ng/mL irisin for 7days. mRNA enrichment from 100 ng of total purified RNA and Illumina sequencing libraries preparation was performed using Kapa stranded mRNA Hyper Prep (Roche Sequencing Solutions, Pleasanton, CA, USA). Gene libraries were multiplexed in an equimolar pool and were sequenced on an Illumina NextSeq 500 with single-end 75 bp reads. Raw reads were aligned to the UCSC mm10 reference genome using a STAR aligner (version STAR_2.4.2a), and raw gene counts were quantified using the quantMode GeneCounts flag. Differential expression testing was performed using Limma and DESeq2. RNAseq analysis was performed using the VIPER snakemake pipeline. See linked manuscript for full details.