RNA-sequencing of stably engrafted miR-99 KD LSKs

We FACS-purified GFP+ LSK cells from mice transplanted with stably engrafted miR-99 KD and Scr HSCs and performed RNA-sequencing, demonstrating miR-99 KD results in significant depletion of hematopoietic stem cell gene expression signature and induces a differentiated progenitor gene expression signature. FACS purification of Scr and miR-99 KD LSKs from 2 mice in each group

我们通过荧光激活细胞分选（Fluorescence-Activated Cell Sorting, FACS），从移植了稳定嵌合miR-99敲低（miR-99 KD）与乱序对照（Scramble, Scr）造血干细胞（Hematopoietic Stem Cells, HSCs）的小鼠中纯化得到绿色荧光蛋白阳性（Green Fluorescent Protein-positive, GFP+）LSK细胞，并开展RNA测序（RNA-sequencing）。实验结果表明，miR-99 KD可显著降低造血干细胞基因表达特征的丰度，并诱导分化型祖细胞的基因表达特征。本研究从每组2只小鼠中分别分选得到Scr与miR-99 KD LSK细胞。