Journal: Immunity

Article Title: TCF-1-centered transcriptional network drives an effector versus exhausted CD8 T cell fate decision

doi: 10.1016/j.immuni.2019.09.013

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: Retroviral vector (RV) experiments The TCF-1 p33 (MR226713) cDNA clone was obtained from OriGene and the TCF-1 p45 cDNA clone was extended from the TCF-1 p33 cDNA using PCR.

Techniques: Flow Cytometry, Staining, In Vitro, Transduction, Recombinant, Cell Isolation, Transfection, Construct, Over Expression, Plasmid Preparation, RNA Sequencing Assay, Multiplex Assay, SYBR Green Assay, Western Blot, Immunofluorescence, Software

Journal: Cell Death & Disease

Article Title: Hippo kinases Mst1 and Mst2 maintain NK cell homeostasis by orchestrating metabolic state and transcriptional activity

doi: 10.1038/s41419-024-06828-x

Figure Lengend Snippet: A Heatmap analysis of RNA-seq data reveals differential expression of certain transcription factors between NK cells from Mst1/2 fl/fl and Mst1/2 fl/fl CD122 Cre mice. Flow cytometry analysis ( B ) and MFI intensity ( C ) demonstrate the levels of Eomes, T-bet, E4BP4 and TCF1 in splenic NK cells from Mst1/2 fl/fl and Mst1/2 fl/fl CD122 Cre mice ( n ≥ 4). D Gene-set enrichment analysis of RNA-seq data indicates a negative enrichment of TCF1-activated genes in Mst1/2-deficient NK cells compared to WT NK cells. E Flow cytometry analysis (left) and MFI intensity (right) reveal the levels of TCF1 in NK cells from the spleen of Mst1/2 fl/fl and Mst1/2 fl/fl CD122 Cre mice upon stimulation with or without IL-15 (10 ng/mL) for 16 h ( n = 4). F – I Representative plots (left panel) and summary data (right panel) illustrate the percentage of NK cells ( F ), the MFI of total cellular ROS levels ( G ), the percentage of 7AAD + NK cells (H) and distribution pattern of different NK cell subsets (I) in the spleen of BM chimeric mice ( n ≥ 3). Data B , C and E are representative of three independent experiments with similar results. Data F – I are representative of two independent experiments with similar results.

Article Snippet: Monoclonal antibodies against mouse TCF1 (Cat#9066S), phospho-Mob1 (Thr35) (Cat#8699), Bim (C34C5) Rabbit mAb (Cat#12186), Phospho-Stat5 (Tyr694) (Cat#9359), phospho-p44/42 MAPK (Erk1/2) (Thr202/Tyr204) (Cat#4370S) and polyclonal antibodies against mouse Mst1 (Cat#3682) were obtained from Cell Signaling Technology (Beverly, MA).

Techniques: RNA Sequencing, Quantitative Proteomics, Flow Cytometry

Journal: Cell Death & Disease

Article Title: Hippo kinases Mst1 and Mst2 maintain NK cell homeostasis by orchestrating metabolic state and transcriptional activity

doi: 10.1038/s41419-024-06828-x

Figure Lengend Snippet: A , B Flow cytometry analysis (left) and MFI intensity (right) demonstrate the expression levels of p-STAT5, p-S6, p-AKT473 and p-Erk in NK cells from Mst1/2 fl/fl and Mst1/2 fl/fl CD122 Cre mice upon stimulation with or without IL-15 (50 ng/mL) for 30 min ( n ≥ 4). C Gene-set enrichment analysis of RNA-seq data shows the enrichment of hallmark-IL6-JAK-STAT3-signaling related genes in Mst1/2-deficient NK cells compared to WT NK cells. D Flow cytometry analysis (left) and MFI intensity (right) reveals the expression level of p-STAT3 (Ser727) in NK cells from Mst1/2 fl/fl and Mst1/2 fl/fl CD122 Cre mice upon stimulation with or without IL-15 (50 ng/mL) for 30 min ( n = 4). E ChIP-qPCR analysis demonstrated binding of p-STAT3 conserved motifs in the Tcf7 5′ regulatory region (−17.5 kb), while no binding was observed in a region without p-STAT3-binding motifs (+0.2 kb), serving as a negative control. The experiments were using sorted NK cells from WT mice with anti-p-STAT3 antibody or isotype-matched IgG. F – I WT splenocytes were treated with DMSO, Mst1 inhibitor XMU-MP-1 (1 mM, MedChemExpress) or p-STAT3 inhibitor Stattic (1 mM, MedChemExpress) for 24 h, followed by flow cytometry analysis. The representative plots (left) and summary data (right) show the expression level of p-STAT3 (F), TCF1 ( G ), total cellular ROS ( H ) and the percentage of 7AAD + NK cells ( I ) ( n = 4). J Schematic illustration of Mst1/2 regulate NK cell survival and function via controlling metabolic state and transcriptional activity. Data A , B and D – I are representative of three independent experiments with similar results.

Article Snippet: Monoclonal antibodies against mouse TCF1 (Cat#9066S), phospho-Mob1 (Thr35) (Cat#8699), Bim (C34C5) Rabbit mAb (Cat#12186), Phospho-Stat5 (Tyr694) (Cat#9359), phospho-p44/42 MAPK (Erk1/2) (Thr202/Tyr204) (Cat#4370S) and polyclonal antibodies against mouse Mst1 (Cat#3682) were obtained from Cell Signaling Technology (Beverly, MA).

Techniques: Flow Cytometry, Expressing, RNA Sequencing, ChIP-qPCR, Binding Assay, Negative Control, Activity Assay

Journal: Immunity

Article Title: TCR-independent CD137 (4–1BB) signaling promotes CD8 + -exhausted T cell proliferation and terminal differentiation

doi: 10.1016/j.immuni.2023.06.007

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: Anti-human/mouse Tcf1/tcf7 AF488 [C63D9] , Cell Signaling Technology , Cat#: 6444S.

Techniques: Purification, Recombinant, Lysis, Negative Control, Cell Isolation, Staining, Knock-Out, Isolation, Control, Infection, RNA Sequencing, Software, Gene Expression