Project IDOEP004811
Project NameLoss of Setd2 induces systemic inflammation via embryo-derived Kupffer-like cell differentiation to drive malignant transformation
Experiment TypeTranscriptomic single cell
ProtocolBM cells separated from old WT and Setd2-MDS mice were stained with biotin-c-Kit antibody on ice for 20 minutes, then washed with MojoSort™ Buffer (480017, BioLegend) at 3,000rpm for 5 minutes. Subsequently, samples labelled with biotin-c-Kit antibody were isolated through MACS assay, then the c-Kit+ BM cells were purified with BD FACSAria™ III. After purification, the BD Rhapsody system (BD Biosciences) was used to capture the transcriptomic information of single cells. Then, a limiting dilution approach was performed to randomly sediment and capture the single cell. Beads with oligonucleotide barcodes were then added to reach a saturated state, ensuring that each cell can be paired with one bead. Cells subjected with beads were captured and used for scRNA-seq. Whole transcriptome libraries were constructed following the BD Rhapsody single-cell whole-transcriptome amplification workflow. Single-cell libraries were quantified using a High Sensitivity DNA chip (Agilent) on a Bioanalyzer 2200 and the Qubit High Sensitivity DNA Assay Kit (Thermo Fisher Scientific). The library was sequenced by HiSeq Xten (Illumina) on a 150 bp paired-end run.
OEX027261
Project IDOEP004811
Project NameLoss of Setd2 induces systemic inflammation via embryo-derived Kupffer-like cell differentiation to drive malignant transformation
Experiment TypeTranscriptomic
ProtocolA total of 1×105 BM LSKs from WT and KO-MDS mice were sorted, washed in DPBS, and lysed in 50 μL lysis buffer (10 mM Tris-HCl pH 7.4, 10 mM NaCl, 3 mM MgCl2, 0.1% (v/v) Igepal CA-630) for the follow-up process, which was performed by TruePrepTM DNA Library Prep Kit V2 for Illumina (TD501, Vazyme) according to the manufacturer’s instructions. Subsequently, the library fragments were purified by VAHTS DNA clean beads (Vazyme, N411-01) for future sequencing according to standard illumina HiSeq/NextSeq platform manuscripts.
OEX027259
Project IDOEP004177
Project NameOEP004177
Experiment TypeTranscriptomic single cell
OEX027323
Project IDOEP004177
Project NameOEP004177
Experiment TypeTranscriptomic single cell
OEX027324
Project IDOEP004177
Project NameOEP004177
Experiment TypeTranscriptomic single cell
OEX024220
Project IDOEP004177
Project NameOEP004177
Experiment TypeTranscriptomic single cell
OEX024221
Project IDOEP004177
Project NameOEP004177
Experiment TypeTranscriptomic single cell
OEX024222
Project IDOEP004177
Project NameOEP004177
Experiment TypeTranscriptomic single cell
OEX024223
Project IDOEP004193
Project NameOEP004193
Experiment TypeGenomic
OEX024236
Project IDOEP004193
Project NameOEP004193
Experiment TypeGenomic
OEX024235