Loss of KDM4B Exacerbates Bone-Fat Imbalance and Mesenchymal Stem Cell Exhaustion in Skeletal Aging [ChIP-seq]

Mesenchymal stem/stromal cells (MSCs) are multipotent progenitor cells with multilineage differentiation potentials. We previously reported histone demethylases KDM4B promoted osteogenesis and inhibited adipogenesis of human MSCs. To identify genome-wide enrichments of KDM4B and the changes in H3K9me3 marks when loss of KDM4B, Primary MSCs were isolated from Prx1Cre; Kdm4bf/f mice and control mice. ChIP-seq assay was performed using KDM4B, H3K9me3 and H3 antibody, respectively. Overall design: Identification of genome-wide enrichments of KDM4B and H3K9me3 marks in mouse bone marrow-derived MSCs. Primary mMSCs were isolated from Prx1Cre; Kdm4bf/f mice (KO) and Prx1Cre; Kdm4bw/w mice (WT). ChIP assay was performed using KDM4B, H3K9me3 and H3 antibody (rabbit anti-KDM4B (1:400, Bethyl Laboratories, Cat No: A301-478A, Lot No.: 2); rabbit anti-KDM4B (1:400, Abcam, Cat No: ab191434, Lot No.: EPR18603-2); rabbit anti-H3K9me3 (1:500, Abcam, Cat No: ab8898, Lot No.: GR3176466-6) and rabbit anti-H3 (1:200, Cell Signaling Technology, Cat#2650, Lot No.: 20)), , respectively. Yeast chromatin was added as a spike-in control for H3K9me3 ChIP-seq. Libraries were prepared using KAPA Hyper Prep Kit (Kapa Biosystems, KK8502) per the manufacturer's protocol. The sequencing was completed using HiSeq 3000 system (Illumina) or Nextseq 500 at the Technology Center for Genomics & Bioinformatics (TCGB) of UCLA.