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...糖皮质激素过量可通过减少骨形成、增加骨吸收引发骨丢失；miR-433-3p是一种微小RNA（microRNA, miRNA），可通过靶向Runt相关转录因子2（Runx2）以及参与Wnt信号通路、蛋白激酶A（protein kinase A, PKA）和内源性糖皮质激素信号通路的RNA分子，在雄性小鼠中负调控骨形成。为探究miR-433-3p对糖皮质激素介导骨丢失的影响，研究人员对在成骨细胞谱系中表达miR-433-3p强效诱饵竞争性抑制剂的转基因小鼠，通过缓释丸剂给予泼尼松龙。与miR-433-3p强效诱饵小鼠相比，经泼尼松龙处理的对照组小鼠骨丢失更为显著，原因在于对照组的破骨细胞活性更高。在完整股骨中，泼尼松龙处理的对照组小鼠的Rankl表达量显著高于miR-433-3p强效诱饵小鼠。令人意外的是，Wnt信号通路负调控因子骨硬化蛋白（Sost）和Dickkopf相关蛋白1（Dkk1）在miR-433-3p强效诱饵小鼠中表达水平更高，且不受泼尼松龙调控。荧光素酶3'UTR报告基因实验证实，Sost是一种新型…… 补充表1：实时定量聚合酶链反应（qRT-PCR）引物集及小鼠Sost 3'UTR克隆引物 补充图1：Col1a1miR-433TuD不在破骨细胞谱系中表达：将原代骨髓单核细胞用巨噬细胞集落刺激因子（M-CSF）+核因子κB受体活化因子配体（RANKL）（各30 ng/ml）培养4天后，叠加tdTomato荧光信号的微分干涉差图像。 ——本数据集为论文“下调成骨细胞谱系中miR-433-3p活性可减弱糖皮质激素介导的骨丢失”的补充数据 https://doi.org/10.5061/dryad.1g1jwsv6s ## 数据与文件结构描述 为探究miR-433-3p对糖皮质激素介导骨丢失的影响，本研究对在成骨细胞谱系中表达miR-433-3p强效诱饵竞争性抑制剂的转基因小鼠，通过缓释丸剂给予泼尼松龙。研究采用组织形态计量学、显微计算机断层扫描（microCT）、组织学、血清学及RNA分析等手段，检测糖皮质激素对骨结构、骨形成与骨吸收参数以及基因表达的影响。 为确定miR-433强效诱饵是否在破骨细胞谱系中表达，研究人员将对照组或miR-433强效诱饵小鼠的原代骨髓单核细胞用M-CSF+RANKL（各30 ng/ml）培养4天，随后对tdTomato荧光进行成像——tdTomato荧光信号可指示强效诱饵的表达情况。作为阳性……