RNA sequencing LSK cells after Cxxc5 knockdown. RNA sequencing LSK cells after Cxxc5 knockdown

To study the effect of knockdown of Cxxc5 on stem/progenitor cells, we used retrovirus to knockdown Cxxc5 expression in the LSK cells. We used this as a surrogate to study the function of Cxxc5 in mouse/stem progenitor cells. Overall design: Stem/progenitor cells enriched bone marrow cells from BALB/c mice were transduced with retrovirus to knockdown Cxxc5. Retrovirus that does not target any mouse genes was used as a control. Infected LSK cells (GFP positive LSK) were flow-sorted. Sorted infected LSK cells from multiple animals were pooled together for RNA isolation.

为探究Cxxc5基因敲低对干/祖细胞的影响，我们采用逆转录病毒在LSK细胞中敲低Cxxc5的表达，并以此作为替代模型，研究Cxxc5在小鼠干/祖细胞中的功能。实验设计：从BALB/c小鼠骨髓中富集得到干/祖细胞，通过逆转录病毒介导Cxxc5基因敲低；以不靶向任何小鼠基因的逆转录病毒作为阴性对照。感染后的LSK细胞（GFP阳性LSK细胞）经流式细胞分选纯化，将多只动物来源的分选后感染LSK细胞混合，用于总RNA提取。