General Information
- Sample ID
- OES354766
- Sample Name
- Old_RNA-seq
- Description
- EII (Cd45- Ter119+ Cd71+), Macrophage (SSClowCd45+Ly6g-Ly6c-F4/80+), neutrophil (Cd45+Ly6c-Ly6g+Cd11b+), monocyte (Cd45+Ly6g-Cd11b+Ly6c+), LSK (Lin- c-Kit+ Sca1+) and liver cell from WT and Setd2-KO MDS mice were used for RNA sequencing.
- Subject type
- Animalia
Sample information
- Organism
- Mus musculus
- Tissue
- bone marrow, liver
- Protocol
- For mature immune cell sorting, BM cells were stained with fluorochrome-labeled antibodies against Cd45, Cd11b, Ly6c, Ly6g and F4/80, and then neutrophil (Cd45+Ly6c-Ly6g+Cd11b+), monocyte (Cd45+Ly6g-Cd11b+Ly6c+), and macrophage (SSClowCd45+Ly6g-Ly6c-F4/80+) cells were sorted on BD FACSAria™ III (BD Biosciences), respectively. For erythroid cell sorting, BM cells were first labeled with biotin-conjugated marker Cd45, and then the magnetic-activated cell sorting (MACS) assay was performed according to the manufacturer’s instructions (76447, BioLegend) to exclude Cd45+ cells. Next, Cd45- cells were incubated with antibodies against Cd45, Cd71 and Ter119 to purify the erythroid precursor (Cd45-Cd71+Ter119+). For HSPC sorting, BM cells were first incubated with five biotin-conjugated lineage antibodies (B220, Ter119, Cd3, Cd11b, and Gr-1), and then using MACS assay to deplete the lineage-positive cells. Lineage-negative cells were subjected to isolate Lin-Sca-1+c-Kit+ (LSK) progenitors by staining with antibodies against c-Kit and Sca-1.
- Projects
-
1
Project ID Project name Submitter Date OEP004811 Loss of Setd2 induces systemic inflammation via embryo-derived Kupffer-like cell differentiation to drive malignant transformation Zhang, Yuanliang 2023-12-07 - Experiments
-
1
Experiment ID Experiment name Experiment type Experiment protocol Date OEX027260 Old_RNA-seq Transcriptomic For bulk RNA-seq, total RNA samples were extracted, and ribosomal RNA (rRNA) was removed using the Ribo-Zero rRNA Removal Kit (MRZMB126, Illumina). Subsequently, rRNA-depleted samples were fragmented, then subjected to first and second strand cDNA synthetization. cDNAs were ligated with sequencing adapters according to standard Novaseq 6000 platform (Illumina) manuscripts. 2024-01-26 - Runs
-
1
Run ID Run Name Project Experiment Sample Data Date OER458862 Old_RNA-seq OEP004811 OEX027260 OES354766 32 2024-01-26
Author Information
- Create Date
- 2024-01-26
- Last Modified
- 2024-01-26
- Submission
- Yuanliang Zhang,Shanghai Institute of Hematology