Project description:To invesitigate the additional paracaspase-independent tumor-promoting effect exist for Malt1. We thus collected E0771-Vector control tumor tissue, E0771 wildtype Malt1 (Malt1-WT) overexpression tumor tissue and E0771 paracaspase-deficient Malt1 (Malt1-PD) overexpression tumor tissue, and sorted out immune cells for 10× Genomics single cell RNA-sequencing (scRNA-seq).
Project description:To reveal the downstream signal change induced by Malt1, we collected mRNA samples from Vector control- and WT-Malt1-expressing E0771 cancer cells, and performed RNA-seq analysis.
Project description:To reveal the downstream paracrine signals from WT-Matl1 or PD-Malt1 tumor cells which responsible for the above observed macrophage phenotypes, we collected mRNA samples from Vector control-, WT-Malt1-, PD-Malt1-, V87R-Malt1-, or L88D-Malt1-expressing E0771 cancer cells, and performed RNA-seq analysis.
Project description:E0771 tumors were implanted either subcutaneously on the left flank or orthotopically in the mammary fat pad and treated with either PBS or FEC + oHSV-1 Mice were sacrificed 5 days after the start of treatment and whole tumour digests were used for RNA extraction
Project description:To understand potential downstream signaling molecules that are responsible for wild type-Malt1 induced resistance of tumor cell to CD8 T cell killing , we first use E0771 cells expressing either -Vector control, wild type-Malt1 (M-WT), or Malt1 PD mutant (M-PD) to coculture with activated CD8 T cell at a effctor: target ratio(E:T ration) of 3 for 12 hours. Then wash the cells with PBS, digest the cells by 0.25% trypsin solution, harvest the cells in 15 ml tubes by centrifugation (1000 rpm 5min) and seed cells into the original plate with DMEM+10%FBS+1%P/S medium. 6 hours after sseding, wash cells with PBS, lyse cells with TRIZOL for RNA extraction, perform reverse transcription and cDNA library for RNA-Seq analysis.
Project description:Major transcriptional changes (290 differentially expressed genes) take place upon downregulation of Lamin A/C in E0771 breast cancer cells
Project description:To investigate the impact of SETD2 mutations on clonal fitness in B actue lymphoblastic leukemia we generated cells lines with SETD KO using CRISPR/Cas9 system. We then performed phenotypic based assays as well as gene expression profiling, mutation burden, and chromtain accessibilty analysis using data obtained from RNA-seq, Whole exom sequencing following drug traetment, and ATAC-seq.
Project description:To study the transcriptome differences of tumor transplanted into WT and KO host, E0771 tumors harvested from WT and KO host were extracted for total RNA and mRNA sequencing was performed. To study the transcriptome differences of WT and KO fat tissue, visceral fat tissue were harvested from obese WT and KO mice and total RNA was extracted and sequenced.
Project description:E-FABP is dominately expressed in macrophages/dendritic cells. Thus E-FABP may play a siginificant role in their immune response to tumor insult. We treated mouse GM-BMMs of different genotype with E0771 breast cancer cells. Then we investigate the global gene expression in these GM-BMMs in response to tumor treatment. Bone marrow cells were collected from naïve C57BL/6 and E-FABP knockout mice of 6~8 week old. GM-BMMs were derived using GM-CSF (in RPMI1640 with 5%FBS) for 7 days before tumor treatment. After 18hr tumor teatment, GM-BMM total RNA was extracted using RNeasy Mini Kit (Qiagen) and subjected to mouse mRNA Gene expression by Affymetrix microarray.
