anti hcd45 pe  (Proteintech)

Journal: bioRxiv

Article Title: TALEN-mediated intron editing of HSPCs enables transgene expression restricted to the myeloid lineage

doi: 10.1101/2024.03.05.583596

Figure Lengend Snippet: Diagram depicting timelines for in vivo experiments. b. Engraftment in blood, bone marrow and spleen in NSG cohorts, measured as the proportion of human cells (hCD45+). c. Engraftment in blood, bone marrow and spleen in SGM3 cohorts, measured as the proportion of human cells (hCD45+). d. Percentage of myeloid, B cell lymphoid, T cell lymphoid and HSPC in the bone marrow of NSG cohorts. e. Percentage of myeloid, B cell lymphoid, T cell lymphoid and HSPC in the bone marrow of SGM3 cohorts. N/A: Not analyzed. Empty circles = SGM3. Filled circles = NSG. Data are represented as mean ± standard deviation. Statistical comparisons were performed using one way ANOVA followed by a Tukey’s Honestly-Significant Difference post-hoc test between each two groups; Tukey’s p values are only shown if ANOVA was significant; ns= non-significant, *p≤0.05, **p≤0.01.

Article Snippet: For neurons and neuroglia from both eGFP-edited and IDUA- edited mouse cohorts, the following antibody panel was used: hCD45-PE-Cy7 (Clone: REA747; Miltenyi), mCD45-APC-Cy7 (Clone: 30-F11; BD Biosciences), and CD11b-PE (Clone: M1/70; Biolegend).

Techniques: In Vivo, Standard Deviation

Journal: bioRxiv

Article Title: TALEN-mediated intron editing of HSPCs enables transgene expression restricted to the myeloid lineage

doi: 10.1101/2024.03.05.583596

Figure Lengend Snippet: Edited HSPCs as a vehicle for therapeutic delivery across the blood-brain barrier. a. Engraftment in brain, measured as the proportion of human cells (hCD45+) in the leukocyte compartment in NSG and SGM3 animals injected with HSPC edited with the eGFP or IDUA cassettes. b. Percentage of CD11b + among the human cells from animals injected with HSPC edited with the eGFP or IDUA cassettes. c. Percentage of eGFP+ cells within the human fraction of cell isolated from the brain of animals engrafted with HSPC edited with eGFP cassettes. d. Percentage of eGFP+ cells within the human CD11b + and CD11b - fractions from the brain of animals engrafted with HSPC edited with eGFP cassettes. e. Percentage of edited alleles among human cells found in the brain of animals engrafted with HSPC edited with IDUA cassettes. Empty circles = SGM3. Filled circles = NSG. Data are represented as mean ± standard deviation. Statistical comparisons were performed using unpaired T tests when comparing two groups , or one way ANOVA when comparing three groups followed by a Tukey’s Honestly- Significant Difference post-hoc test between each two groups ( , 5b). Tukey’s p values are only shown if ANOVA was significant; ns= non-significant, *p≤0.05.

Article Snippet: For neurons and neuroglia from both eGFP-edited and IDUA- edited mouse cohorts, the following antibody panel was used: hCD45-PE-Cy7 (Clone: REA747; Miltenyi), mCD45-APC-Cy7 (Clone: 30-F11; BD Biosciences), and CD11b-PE (Clone: M1/70; Biolegend).

Techniques: Injection, Isolation, Standard Deviation

Journal: Frontiers in immunology

Article Title: Stromal depletion by TALEN-edited universal hypoimmunogenic FAP-CAR T cells enables infiltration and anti-tumor cytotoxicity of tumor antigen-targeted CAR-T immunotherapy.

doi: 10.3389/fimmu.2023.1172681

Figure Lengend Snippet: FIGURE 3 FAP(hF1) UCAR T-cells successfully deplete CAFs and accumulate in TNBC tumor stroma in vivo (A) Schematic of UCAR T-cell treatment and analysis of orthotopic TNBC tumors implanted in NSG mice. (B) Bar graph representing weights of tumor harvested from mice treated with control UT, FAP(hF1) UCAR, FAP(mF3) UCAR or Meso UCAR T-cells as per schematic in (A). Bars show the means ± SEM; P-values determined by Student t test (two-tailed, unpaired), n=2. (C) Flow cytometry plot of human CD45+ UCAR T-cell population in orthotopic TNBC tumors harvested from mice treated as indicated. (D) Box and whiskers plot representing quantitation of total number of CD45+ cells per gram tumor, calculated using (C). P-values determined by Student t test (two-tailed, unpaired), n=2. (E) Stacked bars plot representing CD4+ and CD8+ population, as percentage of total human CD45+ cells, in orthotopic TNBC tumors from mice treated with indicated UCAR T-cells. Bars show the means ± SEM, n=2. (F) Immunohistochemistry for detection of human CD8+ cells in orthotopic TNBC tumors from mice treated with indicated UCAR T-cells. Yellow arrows indicate human CD8 stained cells. (G) Bar graph representing quantitation of (F), each section divided into 4 fields of view. Bars show the means ± SEM; P-values determined by Student t test (two-tailed, unpaired), n=2. (H) Box and whiskers plot representing quantitation of total number of human FAP+ cells per gram tumor, as determined by flow cytometry analysis of orthotopic TNBC tumors from mice treated with indicated UCAR T-cells. P-values determined by Student t test (two-tailed, unpaired), n=2. (I) Trichrome staining of orthotopic TNBC tumor sections from mice treated with indicated UCAR T-cells. Collagen network indicated by blue staining. (J) Bar graph representing quantitation of blue stained Collagen area per section (F), measured using ImageJ. Bars show the means ± SEM; P-values determined by Student t test (two-tailed, unpaired), n=2. *P < 0.05; **P < 0.01; ns, not significant.

Article Snippet: Viability Fixable Viability Dye eFluor 780 Invitrogen 65086514 UCAR T Phenotype FAP Protein Fluorescein (FITC) AcroBiosystems FAP-HF263 Mesothelin protein R-Phycoerythrin (PE) AcroBiosystems MSN-HP2H8 TCRa/b Allophycocyanin (APC) Miltenyi Biotec 130-098-859 HLA-ABC VioBlue Miltenyi Biotec 130-101-459 Tumor analysis Antigen Fluorochrome Company hCD45 R-Phycoerythrin (PE) Miltenyi 130-110-632 mCD45 Brilliant Violet-421 (BV421) BD Biosciences 560501 CD4 Phycoerythrin-Vio770 (PE-Vio770) Miltenyi Biotec 130-113-227 CD8 Brilliant Violet 510 (BV510) BD Bioscience 563919 hFAP Alexa Fluor 488 R&D Systems MAB9727-100 hMesothelin Allophycocyanin (APC) R&D Systems FAB32652P F rontiers in Imm unology

Techniques: In Vivo, Control, Two Tailed Test, Flow Cytometry, Quantitation Assay, Immunohistochemistry, Staining, Cytometry

Journal: Nature cancer

Article Title: P2RY2-AKT activation is a therapeutically actionable consequence of XPO1 inhibition in acute myeloid leukemia

doi: 10.1038/s43018-022-00394-x

Figure Lengend Snippet: a) Confirmation of PDX engraftment by measurement of circulating human CD45+ cells transplanted into NOG-EXL mice (n=6 biologically independent mice per group); data are presented as mean +/− s.d.

Article Snippet: Twelve days after injection, mice were assessed for engraftment: peripheral blood samples and bone marrow biopsies were resuspended in PBS, 0.5% BSA, 2mM EDTA prior to staining with anti-human PE-Vio770-coupled CD45 (hCD45) antibody (130-113-119, Miltenyi).

Techniques: