SRA

To clarify the mechanism through which HEMGN (also known as EDAG in human) controls the survival and functions of hematopoietic stem and progenitor cells (HSPCs) under transplantation stress, we compared the transcriptome of Hemgn-/- and WT HSPCs isolated from primary recipients 6h after transplantation by RNA sequencing (RNA-Seq). Overall design: 2×10^7 bone marrow cells from Hemgn-/- or WT (CD45.2+) mice were injected intravenously via the tail vein into recipients mice (CD45.1+) previously irradiated with a split dose of 9Gy (4.5Gy+4.5Gy). At 6h after transplantation, donor HSPCs (CD45.2+Lin-Sca-1+c-Kit+) were isolated and total RNA was isolated using the QIAGEN RNeasy Micro Kit (QIAGEN). Each group contain three replicates. Smart-Seq2 method was used to produce cDNA. The cDNA production was checked by Qubit® 3.0 Flurometer and Agilent 2100 Bioanalyzer to ensure the expected production with length around 1~2kbp. Then the cDNA was sheared randomly by ultrasonic waves for Illumina library preparation protocol including DNA fragmentation, end repair, 3' ends A-tailing, adapter ligation, PCR amplification and library validation. After library preparation, PerkinElmer LabChip® GX Touch and Step OnePlus™ Real-Time PCR System were introduced for library quality inspection. Qualified libraries were then loaded on Illumina Hiseq platform for PE150 sequencing at the ANNOROAD GENOME Co., Ltd. (Beijing, CN) following the vendor's recommended protocol.