A pan-family screen of Nuclear Receptors in immunocytes reveals ligand-dependent inflammasome control [scRNA-seq]

To examine more closely the varied effects of RA receptor deletions on peritoneal MFs, and determine whether inactivation of RXRα and RARγ elicited superimposable effects on different LPM clusters (as predicted if they operate as a heterodimer, a known propensity of RXR proteins), we performed single-cell RNA sequencing (scRNAseq) analyses on total CD11b+CD115+ peritoneal cells from BMC mice edited for several RA receptors, 17 days after reconstitution. Total CD11b+CD115+ peritoneal macrophages were sorted from BMC mice carrying RA receptor mutants (i.e., RORα, RXRβ ,RARγ and RXRα) and internal controls, 17 days after reconstitution. Each in biological duplicate, and all multiplexed by hashtagging into the same run for optimal comparability.

为更精准地解析视黄酸（RA）受体（Retinoic Acid Receptor）对腹膜巨噬细胞（peritoneal macrophages, 简称MFs）的多元调控效应，并验证RXRα与RARγ的失活是否会对不同LPM细胞簇产生叠加等效的调控效应——若二者以异二聚体形式发挥功能（这是RXR蛋白已知的作用倾向），则该预测成立——我们对经多种视黄酸受体基因编辑的骨髓嵌合小鼠（Bone Marrow Chimeric mice, 简称BMC小鼠）的总CD11b阳性CD115阳性腹膜细胞开展了单细胞RNA测序（single-cell RNA sequencing, 简称scRNAseq）分析，样本采集于造血重建后第17天。
我们从携带视黄酸受体突变体（即RORα、RXRβ、RARγ及RXRα）及内参对照的骨髓嵌合小鼠中，分选得到总CD11b阳性CD115阳性腹膜巨噬细胞，样本采集于造血重建后第17天。
所有样本均设置两份生物学重复，并通过hashtag标记技术进行多重混样，纳入同一测序批次以实现最优的组间可比性。