Gene expression profiling in Tmem30a-/-LSCs

Hematopoietic stem cells (LinlowSca1+c-Kit+) (LSK cells) were sorted from the BM of WT and Tmem30a-/- mice at day 7 post tamoxifen injection using a fluorescence activated cell sorter Aria III (Becton Dickinson) . Total RNA was extracted from LSK cells using the RNeasy Mini kit (Qiagen). RNA samples were reverse transcribed, amplified using the Ovation Pico WTA System V2 (NuGEN Technologies), and biotin labeled with the Encore Biotin Module (NuGEN Technologies). RNA-seq was performed with Illumina MiSeq system. Gene set enrichment analysis was performed as previously described.