Project description:MED23, a subunit of the Mediator coactivator complex, is important for the expression of a subset of MAPK/ERK pathway-dependent target genes; however, the genes in this subset varies between cell types. MAPK/ERK pathway-dependent processes are essential for T-cell development and function, but whether MED23 has a role in this context is unknown. We generated Med23 conditional knockout mice and induced Med23 deletion in early T cell development using the lineage specific Lck-Cre transgene. While the total cell number and distribution of cell populations in the thymuses of Med23flox/flox;Lck-Cre mice were essentially normal, MED23 null T-cells failed to efficiently populate the peripheral lymphoid organs. MED23 null thymocytes displayed decreased expression of the MAPK/ERK-responsive genes Egr1, Egr2, as well as of the membrane glycoprotein Cd52 (CAMPATH-1). MED23 null CD4 single-positive thymocytes also showed decreased expression of KLF2 (LKLF), a T cell master regulatory transcription factor. Indeed, similarities between the phenotypes of mice lacking MED23 or KLF2 in T-cells suggest that KLF2 deficiency in MED23 null T-cells is one of their key defects. Mechanistic experiments using MED23 null MEFs further suggest that MED23 is required for full activity of the MAPK-responsive transcription factor MEF2, which has previously been shown to mediate Klf2 expression. In summary, our data indicate that MED23 has critical roles in enabling T-cells to populate the peripheral lymphoid organs, possibly by potentiating MEF2-dependent expression of the T-cell transcription factor KLF2. 12 samples, 2 of each genotype (Lck-Cre, Med23flox/flox and Med23flox/flox;Lck-Cre) both with mock and anti-CD3 treatment
Project description:tPTEN-/- mice display a deletion of the PTEN tumor suppressor gene specifically in T cells (cross PTEN flox/flox x lck-Cre). They develop T cell lymphoma with a primary thymic tumor and invasion of most organ at late stage of the disease. Gene profiling was performed on whole tumors (ST3 stage 3 invasive lymphoma) and normal thymocytes (wt) using Affymetrix-MoGene-1_0-st-v1 chips.
Project description:Human CD4+ T cells were left untreated or stimulated with 5 μg/ml plate-bound anti-CD3 antibody (clone OKT3, 14-0037-82, eBioscience) and 10 μg/ml soluble anti-CD28 antibody (clone CD28.2, 14-0289-82, eBioscience) for 6 h in RPMI1640 medium supplemented with 10% FBS, 1% L-glutamine, and 1% penicillin-streptomycin.
Project description:We report the differential mRNA expressino of WT and Chi3l1 KO Th1 cells. We cultured WT and Chi3l1 KO naive CD4 T cells under Th1 skewing condition : plate-bound anti-CD3/28 antibody (2ug/mL), IL-12 0.2ng/mL, IL-2 50U/mL, anti-IL-4 neutralizing antibody 2ug/mL, for 3 days. We found different Th1 regulatory and tumoricidal-related gene expression in Chi3l1 KO T cells.
Project description:compare gene expression profiles between normal and anergic T cells and identify upregulated genes in anergic T cells Experiment Overall Design: RNA from normal Th1 T cell clone and anergic Th1 T cell clone made anergic by plate-bound anti-CD3 antibody were isolated and amplified for microarray analysis
Project description:Control (cMet+ T cells less than 0.5%) and cMet-enriched (7%) T cells were activated with plate-bound anti-CD3/CD28 and gene tracription was measured
Project description:Mouse CD4 naïve T cells, isolated from spleens of WT or TLGMNKO mice, were activated with plate-bound anti-mouse CD3 and soluble anti-mouse CD28 for 3 days, and then characterized by 4D label-free proteomic analysis using nano-liquid chromatography-tandem mass spectrometry
Project description:tPTEN-/- mice display a deletion of the PTEN tumor suppressor gene specifically in T cells (cross PTEN flox/flox x lck-Cre). They develop T cell lymphoma with a primary thymic tumor and invasion of most organ at late stage of the disease.
Project description:Steroid receptor coactivator 3 (SRC3) was reported to regulates pathogenic Th17 cell differentiation in vitro and in vivo.However, the role of SRC2, another member of SRC family, and its relationship with SRC3 in the regulation of Th17 cell differentiation remain unknown. We used RNA-seq to study the global function of SRC2 and SRC3. Naive CD4+ T cells were activated with 5ug/ml plate-bound anti-CD3/28, and cultured with TGF-β+IL-6 or with IL-6+IL-1β+IL23. 4days later, cells were restimulated with plate-bound anti-CD3 for 4hrs for RNA-seq.
Project description:Lamtor1-KO BMDCs isolated from Lamtor1flox/flox X CD11c-Cre mice were lysed by Lysis Buffer A and immunoprecipitated by anti-Lamtor1 antibody (D11H6).
