Role of growth plate Nox2 in alcoholic osteopenia

To study the role of Nox2 in ethanol toxicity on the bone growth plate chondrocytes, we generated Nox2 conditional knockout mice (CKO), in which the catalytic subunit of Nox2, Cybb, is deleted in chondrocytes using a Cre-lox system, where Cre is expressed from the Col2a1 promoter. CKO mice and floxed control mice were fed an ethanol-containing Lieber De-Carli-based diet or pair-fed a control diet for 8 weeks starting at 5-6 weeks of age. As both the Nox2 genotype and ethanol diminished the number of chondrocytes in the growth plates, we conducted an RNA-Seq analysis of the growth-plate containing regions of the femurs. Femurs were separated into femoral shaft, femoral marrow and the femoral end pieces containing the growth plates (Pedersen et al. Bone Rep. 2019 May 31;11:100211. doi: 10.1016/j.bonr.2019.100211.). RNA was isolated from femoral end pieces from female mice. Each sample of RNA subjected to RNA-Seq was a pool of RNA from three mice. The design is a 2 X 2 factorial design with two genotypes (CKO and floxed controls) and two diets (ethanol-fed and pair-fed) with three samples per group. The RNA-Seq libraries and initial analyses were performed by the commercial vendor Azenta/GeneWiz.