mRNA expression in in vivo osteocytes as function of PPARG

Description

The skeleton is one of the largest organs in the body, wherein metabolism is integrated with systemic energy metabolism. However, the bioenergetic programming of osteocytes, the most abundant bone cells coordinating bone metabolism, is not well defined. Here, using a mouse model with partial penetration of an osteocyte-specific PPARG deletion, we demonstrate that PPARG controls osteocyte bioenergetics and their contribution to systemic energy metabolism independently of circulating sclerostin levels. Osteocyte RNA from γOTKO (n=4) and Ctrl (n=3) 4 mo old male mice was isolated using TRIZOL and transcriptomic analysis was performed by Arraystar Inc. (Rockville, MD, USA) using the Mouse LncRNA Array v4.0 platform (8 x 60K, Arraystar, Inc). To increase stringency, the same RNA was also analyzed with Next Generation Sequencing (NGS) using flow of NovaSeq SP 100 cycles (Wayne State University Bioinformatics Core, Detroit, MI). In addition, NGS analysis as above was performed on PPARG CRISPR-edited clones created using MLO-Y4 osteocytic cell line.

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Osteocyte RNA from γOTKO (n=4) and Ctrl (n=3) 4 mo old male mice was isolated using TRIZOL and transcriptomic analysis was performed by Arraystar Inc. (Rockville, MD, USA) using the Mouse LncRNA Array v4.0 platform (8 x 60K, Arraystar, Inc). To increase stringency, the same RNA was also analyzed with Next Generation Sequencing (NGS) using flow of NovaSeq SP 100 cycles (Wayne State University Bioinformatics Core, Detroit, MI). Similarly, transcriptomes of γY4KO clones with highest KO scores were analyzed by NGS, as above.

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