Supplemental data for: Decreasing miR-433-3p activity in the osteoblast lineage blunts glucocorticoid-mediated bone loss

Glucocorticoid excess causes bone loss due to decreased bone formation and increased bone resorption; miR-433-3p is a miRNA that negatively regulates bone formation in male mice by targeting Runx2 as well as RNAs involved in Wnt, protein kinase A and endogenous glucocorticoid signaling. To examine the impact of miR-433-3p on glucocorticoid-mediated bone loss, transgenic mice expressing a miR-433-3p tough decoy inhibitor in the osteoblast lineage were administered prednisolone via slow-release pellets. Bone loss was greater in control mice treated with prednisolone compared with miR-433-3p tough decoy mice due to higher osteoclast activity in the controls. In whole femurs, Rankl was significantly higher in prednisolone-treated controls compared with miR-433-3p tough decoy mice. Surprisingly, negative regulators of Wnt signaling Sost and Dkk1 were higher in miR-433-3p tough decoy mice and were unaffected by prednisolone. Luciferase-3’ UTR reporter assays demonstrated that Sost is a novel ..., Supplemental Table 1: primer sets for qRT-PCR and murine Sost 3' UTR cloning
Supplemental Figure 1:The Col1a1miR-433TuD is not expressed in the osteoclast lineage: Differential interference contrast images overlaid by tdTomato signal in primary bone marrow monocytes cultured with MCSF+RANKL (30 ng/ml each) for 4 days., , # Supplemental data for \"Decreasing miR-433-3p activity in the osteoblast lineage blunts glucocorticoid-mediated bone loss\"

[https://doi.org/10.5061/dryad.1g1jwsv6s](https://doi.org/10.5061/dryad.1g1jwsv6s)

## Description of the data and file structure

To examine the impact of miR-433-3p on glucocorticoid-mediated bone loss, transgenic mice expressing a miR-433-3p tough decoy competitive inhibitor in the osteoblast lineage were administered prednisolone via slow-release pellets. Histomorphometry, microCT, histology, serum and RNA analyses were used to examine the impact of the glucocorticoid on bone structure, bone formation and resorption parameters and gene expression.  

To determine whether the miR-433 tough decoy was expressed in the osteoclast lineage, primary bone marrow monocytes from control or miR-433 tough decoy mice were cultured with MCSF+RANKL (30 ng/ml each) for 4 days, and imaged for tdTomato fluorescence, which would signify expression of the tough decoy. As a posit...