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celltrace tm violet cell proliferation kit  (Thermo Fisher)


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    Thermo Fisher celltrace tm violet cell proliferation kit
    (A) Donor H2 Ag in splenic FDCs (inset, arrows) after transplantation of B6 (H2 b ) hearts in BALB/c mice. Original magnification ×200. Scale bars, 20 μm. (B) Quantification with ImageJ of donor H2 Ag spots per FDC network on spleens of BALB/c mice transplanted with B6 hearts. (C) Donor H2 Ag spots per FDC network on BALB/c LNs draining B6 skin allografts assessed using ImageJ. (D) Deep SIM of donor H2 Ag spots in FDCs of a BALB/c LN draining a B6 skin graft. Scale bar, 2 μm. ROI, region of interest. (E and F) 2P-microscopy of FDCs in the spleen of a BALB/c mouse transplanted with a B6 heart releasing RFP-sEVs (E) and a BALB/c LN draining a B6 skin graft releasing RFP-sEVs (F). Arrows: graft-RFP-sEVs on the surface (red) or inside (yellow) of FDCs (green). <t>CellTrace</t> Violet-B cells indicate B cell follicles. FDCs were labeled with AF488-CD21/CD35 Ab. Original magnification ×25. Scale bars, 20 μm. (G) Colocalization by confocal microscopy of donor (B6, IA b + H2K b ) allo-Ags in FDCs of LNs draining CMV Cre/+ LSL -RFP-CD63 B6 skin grafted in BALB/c mice. Scale bar, 10 μm. Mander’s colocalization coefficient of graft-sEVs and donor IA b + H2K b Ab analyzed using Imaris. Each triangle represents one FDC network. In (E) and (F), RFP spots per FDC network z stacks were quantified within 90-μm-thick z stacks using Imaris. Experiments were done on 4 spleens per POD (A and B), 6 graft-dLNs per POD (C), 3 graft-dLNs (D), 4 spleens (E), 4 graft-dLNs (F), and 12 graft-dLNs (G). In (B), (C), (E), and (F), comparisons were made by multiple unpaired two-tailed Student’s test. Error bars, means ± SD; * p < 0.05, ** p < 0.01, *** p < 0.001, and **** p < 0.0001; NS, not significant.
    Celltrace Tm Violet Cell Proliferation Kit, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/celltrace%E2%84%A2+violet+cell+proliferation+kit/celltrace+violet/pmc12281710-105-0-10
    Average 90 stars, based on 1 article reviews
    celltrace tm violet cell proliferation kit - by Bioz Stars, 2026-10
    90/100 stars

    Images

    1) Product Images from "Transplants foster B cell alloimmunity by relaying extracellular vesicles to follicular dendritic cells"

    Article Title: Transplants foster B cell alloimmunity by relaying extracellular vesicles to follicular dendritic cells

    Journal: Cell reports

    doi: 10.1016/j.celrep.2025.115832

    (A) Donor H2 Ag in splenic FDCs (inset, arrows) after transplantation of B6 (H2 b ) hearts in BALB/c mice. Original magnification ×200. Scale bars, 20 μm. (B) Quantification with ImageJ of donor H2 Ag spots per FDC network on spleens of BALB/c mice transplanted with B6 hearts. (C) Donor H2 Ag spots per FDC network on BALB/c LNs draining B6 skin allografts assessed using ImageJ. (D) Deep SIM of donor H2 Ag spots in FDCs of a BALB/c LN draining a B6 skin graft. Scale bar, 2 μm. ROI, region of interest. (E and F) 2P-microscopy of FDCs in the spleen of a BALB/c mouse transplanted with a B6 heart releasing RFP-sEVs (E) and a BALB/c LN draining a B6 skin graft releasing RFP-sEVs (F). Arrows: graft-RFP-sEVs on the surface (red) or inside (yellow) of FDCs (green). CellTrace Violet-B cells indicate B cell follicles. FDCs were labeled with AF488-CD21/CD35 Ab. Original magnification ×25. Scale bars, 20 μm. (G) Colocalization by confocal microscopy of donor (B6, IA b + H2K b ) allo-Ags in FDCs of LNs draining CMV Cre/+ LSL -RFP-CD63 B6 skin grafted in BALB/c mice. Scale bar, 10 μm. Mander’s colocalization coefficient of graft-sEVs and donor IA b + H2K b Ab analyzed using Imaris. Each triangle represents one FDC network. In (E) and (F), RFP spots per FDC network z stacks were quantified within 90-μm-thick z stacks using Imaris. Experiments were done on 4 spleens per POD (A and B), 6 graft-dLNs per POD (C), 3 graft-dLNs (D), 4 spleens (E), 4 graft-dLNs (F), and 12 graft-dLNs (G). In (B), (C), (E), and (F), comparisons were made by multiple unpaired two-tailed Student’s test. Error bars, means ± SD; * p < 0.05, ** p < 0.01, *** p < 0.001, and **** p < 0.0001; NS, not significant.
    Figure Legend Snippet: (A) Donor H2 Ag in splenic FDCs (inset, arrows) after transplantation of B6 (H2 b ) hearts in BALB/c mice. Original magnification ×200. Scale bars, 20 μm. (B) Quantification with ImageJ of donor H2 Ag spots per FDC network on spleens of BALB/c mice transplanted with B6 hearts. (C) Donor H2 Ag spots per FDC network on BALB/c LNs draining B6 skin allografts assessed using ImageJ. (D) Deep SIM of donor H2 Ag spots in FDCs of a BALB/c LN draining a B6 skin graft. Scale bar, 2 μm. ROI, region of interest. (E and F) 2P-microscopy of FDCs in the spleen of a BALB/c mouse transplanted with a B6 heart releasing RFP-sEVs (E) and a BALB/c LN draining a B6 skin graft releasing RFP-sEVs (F). Arrows: graft-RFP-sEVs on the surface (red) or inside (yellow) of FDCs (green). CellTrace Violet-B cells indicate B cell follicles. FDCs were labeled with AF488-CD21/CD35 Ab. Original magnification ×25. Scale bars, 20 μm. (G) Colocalization by confocal microscopy of donor (B6, IA b + H2K b ) allo-Ags in FDCs of LNs draining CMV Cre/+ LSL -RFP-CD63 B6 skin grafted in BALB/c mice. Scale bar, 10 μm. Mander’s colocalization coefficient of graft-sEVs and donor IA b + H2K b Ab analyzed using Imaris. Each triangle represents one FDC network. In (E) and (F), RFP spots per FDC network z stacks were quantified within 90-μm-thick z stacks using Imaris. Experiments were done on 4 spleens per POD (A and B), 6 graft-dLNs per POD (C), 3 graft-dLNs (D), 4 spleens (E), 4 graft-dLNs (F), and 12 graft-dLNs (G). In (B), (C), (E), and (F), comparisons were made by multiple unpaired two-tailed Student’s test. Error bars, means ± SD; * p < 0.05, ** p < 0.01, *** p < 0.001, and **** p < 0.0001; NS, not significant.

    Techniques Used: Transplantation Assay, Microscopy, Labeling, Confocal Microscopy, Two Tailed Test

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    Cell Culture:

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    Article Snippet: 384 Suppression assay in vitro 385 For the proliferation assay, PBMCs were labeled with 5μM Cell Trace Violet 386 (CellTraceTM Violet Cell Proliferation kit; Invitrogen, USA) and coculture with 387 CD19+CD1c+ or CD19+CD1c- treated by CpG ODN 2006, db-cAMP and CHIR-99021.

    Article Title: SHMT2 regulates CD8+ T cell senescence via the reactive oxygen species axis in HIV-1 infected patients on antiretroviral therapy
    Article Snippet: The cells were then cultured under the Dynabeads human T-activator CD3/CD28 stimulation for 72 h. For the glycine supplementation experiment, PBMCs from HIV-infected individuals were labelled with the CellTraceTM Violet Cell Proliferation Kit (Thermo Fisher).

    Article Title: Longitudinal analysis of B- and T-cell responses to SARS-CoV-2 recombinant S-protein vaccine S-268019-b in phase 1/2 priming and booster study
    Article Snippet: PBMCs were then stained with CellTraceTM Violet Cell Proliferation Kit (ThermoFisher) in accordance with the manufacturer’s instruction.

    Article Title: SHMT2 regulates CD8+ T cell senescence via the reactive oxygen species axis in HIV-1 infected patients on antiretroviral therapy
    Article Snippet: Following the manufacturer’s instructions, cells were labelled using the CellTraceTM Violet Cell Proliferation Kit (Thermo Fisher, USA, Cat#C34557) 24 h after electroporation.

    Article Title: Dissecting Immunological Mechanisms Underlying Influenza Viral Nucleoprotein-induced Mucosal Immunity Against Diverse Viral Strains.
    Article Snippet: After passing each sample through a 70 μm cell strainer, the cells were labeled using the CellTraceTM Violet Cell Proliferation Kit (Invitrogen, C34571) at a concentration of 1.0 × 106 cells/mL for 20 min at room temperature, protected from light.

    Article Title: Dissecting immunological mechanisms underlying influenza viral nucleoprotein-induced mucosal immunity against diverse viral strains
    Article Snippet: After passing each sample through a 70 μm cell strainer, the cells were labelled using the CellTraceTM Violet Cell Proliferation Kit (Invitrogen, C34571) at a concentration of 1.0 × 106 cells/mL for 20 min at room temperature, protected from light.

    Article Title: DNA demethylation triggers cell free DNA release in colorectal cancer cells.
    Article Snippet: A total of 2.5 × 106 living cells were labeled using the CellTraceTM Violet Cell Proliferation Kit (Thermo Fisher Scientific, C34554), according to the manufacturer’s instructions.

    In Vitro:

    Article Title: Heightened innate immune state induced by viral vector leads to enhanced response to challenge and prolongs malaria vaccine protection
    Article Snippet: CellTraceTM Violet Cell Proliferation Kit , Invitrogen , Cat #C34571.

    Article Title: Ex vivo-generated human CD1c + regulatory B cells by a chemically defined system suppress immune responses and alleviate graft-versus-host disease.
    Article Snippet: 384 Suppression assay in vitro 385 For the proliferation assay, PBMCs were labeled with 5μM Cell Trace Violet 386 (CellTraceTM Violet Cell Proliferation kit; Invitrogen, USA) and coculture with 387 CD19+CD1c+ or CD19+CD1c- treated by CpG ODN 2006, db-cAMP and CHIR-99021.

    Article Title: SHMT2 regulates CD8+ T cell senescence via the reactive oxygen species axis in HIV-1 infected patients on antiretroviral therapy
    Article Snippet: The cells were then cultured under the Dynabeads human T-activator CD3/CD28 stimulation for 72 h. For the glycine supplementation experiment, PBMCs from HIV-infected individuals were labelled with the CellTraceTM Violet Cell Proliferation Kit (Thermo Fisher).

    Article Title: Longitudinal analysis of B- and T-cell responses to SARS-CoV-2 recombinant S-protein vaccine S-268019-b in phase 1/2 priming and booster study
    Article Snippet: PBMCs were then stained with CellTraceTM Violet Cell Proliferation Kit (ThermoFisher) in accordance with the manufacturer’s instruction.

    Article Title: SHMT2 regulates CD8+ T cell senescence via the reactive oxygen species axis in HIV-1 infected patients on antiretroviral therapy
    Article Snippet: Following the manufacturer’s instructions, cells were labelled using the CellTraceTM Violet Cell Proliferation Kit (Thermo Fisher, USA, Cat#C34557) 24 h after electroporation.

    Article Title: Dissecting Immunological Mechanisms Underlying Influenza Viral Nucleoprotein-induced Mucosal Immunity Against Diverse Viral Strains.
    Article Snippet: After passing each sample through a 70 μm cell strainer, the cells were labeled using the CellTraceTM Violet Cell Proliferation Kit (Invitrogen, C34571) at a concentration of 1.0 × 106 cells/mL for 20 min at room temperature, protected from light.

    Article Title: Dissecting immunological mechanisms underlying influenza viral nucleoprotein-induced mucosal immunity against diverse viral strains
    Article Snippet: After passing each sample through a 70 μm cell strainer, the cells were labelled using the CellTraceTM Violet Cell Proliferation Kit (Invitrogen, C34571) at a concentration of 1.0 × 106 cells/mL for 20 min at room temperature, protected from light.

    Article Title: DNA demethylation triggers cell free DNA release in colorectal cancer cells.
    Article Snippet: A total of 2.5 × 106 living cells were labeled using the CellTraceTM Violet Cell Proliferation Kit (Thermo Fisher Scientific, C34554), according to the manufacturer’s instructions.

    Proliferation Assay:

    Article Title: Heightened innate immune state induced by viral vector leads to enhanced response to challenge and prolongs malaria vaccine protection
    Article Snippet: CellTraceTM Violet Cell Proliferation Kit , Invitrogen , Cat #C34571.

    Article Title: Ex vivo-generated human CD1c + regulatory B cells by a chemically defined system suppress immune responses and alleviate graft-versus-host disease.
    Article Snippet: 384 Suppression assay in vitro 385 For the proliferation assay, PBMCs were labeled with 5μM Cell Trace Violet 386 (CellTraceTM Violet Cell Proliferation kit; Invitrogen, USA) and coculture with 387 CD19+CD1c+ or CD19+CD1c- treated by CpG ODN 2006, db-cAMP and CHIR-99021.

    Article Title: SHMT2 regulates CD8+ T cell senescence via the reactive oxygen species axis in HIV-1 infected patients on antiretroviral therapy
    Article Snippet: The cells were then cultured under the Dynabeads human T-activator CD3/CD28 stimulation for 72 h. For the glycine supplementation experiment, PBMCs from HIV-infected individuals were labelled with the CellTraceTM Violet Cell Proliferation Kit (Thermo Fisher).

    Article Title: Longitudinal analysis of B- and T-cell responses to SARS-CoV-2 recombinant S-protein vaccine S-268019-b in phase 1/2 priming and booster study
    Article Snippet: PBMCs were then stained with CellTraceTM Violet Cell Proliferation Kit (ThermoFisher) in accordance with the manufacturer’s instruction.

    Article Title: SHMT2 regulates CD8+ T cell senescence via the reactive oxygen species axis in HIV-1 infected patients on antiretroviral therapy
    Article Snippet: Following the manufacturer’s instructions, cells were labelled using the CellTraceTM Violet Cell Proliferation Kit (Thermo Fisher, USA, Cat#C34557) 24 h after electroporation.

    Article Title: Dissecting Immunological Mechanisms Underlying Influenza Viral Nucleoprotein-induced Mucosal Immunity Against Diverse Viral Strains.
    Article Snippet: After passing each sample through a 70 μm cell strainer, the cells were labeled using the CellTraceTM Violet Cell Proliferation Kit (Invitrogen, C34571) at a concentration of 1.0 × 106 cells/mL for 20 min at room temperature, protected from light.

    Article Title: Dissecting immunological mechanisms underlying influenza viral nucleoprotein-induced mucosal immunity against diverse viral strains
    Article Snippet: After passing each sample through a 70 μm cell strainer, the cells were labelled using the CellTraceTM Violet Cell Proliferation Kit (Invitrogen, C34571) at a concentration of 1.0 × 106 cells/mL for 20 min at room temperature, protected from light.

    Article Title: DNA demethylation triggers cell free DNA release in colorectal cancer cells.
    Article Snippet: A total of 2.5 × 106 living cells were labeled using the CellTraceTM Violet Cell Proliferation Kit (Thermo Fisher Scientific, C34554), according to the manufacturer’s instructions.



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    Thermo Fisher celltrace violet cell proliferation kit
    (A) Donor H2 Ag in splenic FDCs (inset, arrows) after transplantation of B6 (H2 b ) hearts in BALB/c mice. Original magnification ×200. Scale bars, 20 μm. (B) Quantification with ImageJ of donor H2 Ag spots per FDC network on spleens of BALB/c mice transplanted with B6 hearts. (C) Donor H2 Ag spots per FDC network on BALB/c LNs draining B6 skin allografts assessed using ImageJ. (D) Deep SIM of donor H2 Ag spots in FDCs of a BALB/c LN draining a B6 skin graft. Scale bar, 2 μm. ROI, region of interest. (E and F) 2P-microscopy of FDCs in the spleen of a BALB/c mouse transplanted with a B6 heart releasing RFP-sEVs (E) and a BALB/c LN draining a B6 skin graft releasing RFP-sEVs (F). Arrows: graft-RFP-sEVs on the surface (red) or inside (yellow) of FDCs (green). <t>CellTrace</t> Violet-B cells indicate B cell follicles. FDCs were labeled with AF488-CD21/CD35 Ab. Original magnification ×25. Scale bars, 20 μm. (G) Colocalization by confocal microscopy of donor (B6, IA b + H2K b ) allo-Ags in FDCs of LNs draining CMV Cre/+ LSL -RFP-CD63 B6 skin grafted in BALB/c mice. Scale bar, 10 μm. Mander’s colocalization coefficient of graft-sEVs and donor IA b + H2K b Ab analyzed using Imaris. Each triangle represents one FDC network. In (E) and (F), RFP spots per FDC network z stacks were quantified within 90-μm-thick z stacks using Imaris. Experiments were done on 4 spleens per POD (A and B), 6 graft-dLNs per POD (C), 3 graft-dLNs (D), 4 spleens (E), 4 graft-dLNs (F), and 12 graft-dLNs (G). In (B), (C), (E), and (F), comparisons were made by multiple unpaired two-tailed Student’s test. Error bars, means ± SD; * p < 0.05, ** p < 0.01, *** p < 0.001, and **** p < 0.0001; NS, not significant.
    Celltrace Violet Cell Proliferation Kit, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Selective expansion and differentiation of T cells by AP-EVs-Th1 and AP-EVs-Th2 in vitro . (a) Lymph node T cells from C57BL/6 and OT-II transgenic mice were combined in a 1:1 ratio. The combined cells were labeled with <t>CellTrace</t> Violet <t>(CTV)</t> and treated with control EVs or AP-EVs-Th1. (b) The combined cells were co-cultured with AP-EVs-Th1, control EVs, or anti-mouse CD3/CD28 conjugated beads for three days, and <t>proliferation</t> and differentiation of OT-II T cells were analyzed via flow cytometry. (c) Flow cytometric plots of CTV-labeled OT-II (CD45.2/CD45.1) and polyclonal (CD45.1) T cells cultured with anti-mouse CD3/CD28 conjugated beads, AP-EVs-Th1, or control EVs. (d) Percentage of T-bet-expressing CD4 + T cells. (e) Bar graph showing the percentage of T-bet-expressing wild-type (WT) CD4 + T or OT-II T cells under different concentrations of control EVs (black) and AP-EVs-Th1 (orange). (f) The combined cells were co-cultured with AP-EVs-Th2 or control EVs for three days, and the proliferation and differentiation of OT-II T cells were analyzed via flow cytometry. (g) Flow cytometric plots of CTV-labeled OT-II (CD45.2) and polyclonal (CD45.1) T cells cultured with anti-mouse CD3/CD28 conjugated beads, AP-EVs-Th2, or control EVs. (h) Percentage of GATA-3-expressing CD4 + T cells. (i) Bar graph showing the percentage of GATA-3-expressing WT CD4 + or OT-II T cells under different concentrations of control EVs (black) and AP-EVs-Th2 (orange).
    Celltrace Violet (Ctv) Cell Proliferation Kit, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/celltrace%E2%84%A2+violet+cell+proliferation+kit/celltrace+violet/pmc12168395-73-16-22
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    Selective expansion and differentiation of T cells by AP-EVs-Th1 and AP-EVs-Th2 in vitro . (a) Lymph node T cells from C57BL/6 and OT-II transgenic mice were combined in a 1:1 ratio. The combined cells were labeled with <t>CellTrace</t> Violet <t>(CTV)</t> and treated with control EVs or AP-EVs-Th1. (b) The combined cells were co-cultured with AP-EVs-Th1, control EVs, or anti-mouse CD3/CD28 conjugated beads for three days, and <t>proliferation</t> and differentiation of OT-II T cells were analyzed via flow cytometry. (c) Flow cytometric plots of CTV-labeled OT-II (CD45.2/CD45.1) and polyclonal (CD45.1) T cells cultured with anti-mouse CD3/CD28 conjugated beads, AP-EVs-Th1, or control EVs. (d) Percentage of T-bet-expressing CD4 + T cells. (e) Bar graph showing the percentage of T-bet-expressing wild-type (WT) CD4 + T or OT-II T cells under different concentrations of control EVs (black) and AP-EVs-Th1 (orange). (f) The combined cells were co-cultured with AP-EVs-Th2 or control EVs for three days, and the proliferation and differentiation of OT-II T cells were analyzed via flow cytometry. (g) Flow cytometric plots of CTV-labeled OT-II (CD45.2) and polyclonal (CD45.1) T cells cultured with anti-mouse CD3/CD28 conjugated beads, AP-EVs-Th2, or control EVs. (h) Percentage of GATA-3-expressing CD4 + T cells. (i) Bar graph showing the percentage of GATA-3-expressing WT CD4 + or OT-II T cells under different concentrations of control EVs (black) and AP-EVs-Th2 (orange).
    Celltrace Violet Cell Proliferation Kit, For Flow Cytometry, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Selective expansion and differentiation of T cells by AP-EVs-Th1 and AP-EVs-Th2 in vitro . (a) Lymph node T cells from C57BL/6 and OT-II transgenic mice were combined in a 1:1 ratio. The combined cells were labeled with <t>CellTrace</t> Violet <t>(CTV)</t> and treated with control EVs or AP-EVs-Th1. (b) The combined cells were co-cultured with AP-EVs-Th1, control EVs, or anti-mouse CD3/CD28 conjugated beads for three days, and <t>proliferation</t> and differentiation of OT-II T cells were analyzed via flow cytometry. (c) Flow cytometric plots of CTV-labeled OT-II (CD45.2/CD45.1) and polyclonal (CD45.1) T cells cultured with anti-mouse CD3/CD28 conjugated beads, AP-EVs-Th1, or control EVs. (d) Percentage of T-bet-expressing CD4 + T cells. (e) Bar graph showing the percentage of T-bet-expressing wild-type (WT) CD4 + T or OT-II T cells under different concentrations of control EVs (black) and AP-EVs-Th1 (orange). (f) The combined cells were co-cultured with AP-EVs-Th2 or control EVs for three days, and the proliferation and differentiation of OT-II T cells were analyzed via flow cytometry. (g) Flow cytometric plots of CTV-labeled OT-II (CD45.2) and polyclonal (CD45.1) T cells cultured with anti-mouse CD3/CD28 conjugated beads, AP-EVs-Th2, or control EVs. (h) Percentage of GATA-3-expressing CD4 + T cells. (i) Bar graph showing the percentage of GATA-3-expressing WT CD4 + or OT-II T cells under different concentrations of control EVs (black) and AP-EVs-Th2 (orange).
    Celltrace Violet Cell Proliferation Kit 15579992, supplied by Fisher Scientific, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/celltrace%E2%84%A2+violet+cell+proliferation+kit/celltrace+violet+proliferation+dye/bio_rxiv__2025__04__16__649059-277-21-27
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    Image Search Results


    (A) Donor H2 Ag in splenic FDCs (inset, arrows) after transplantation of B6 (H2 b ) hearts in BALB/c mice. Original magnification ×200. Scale bars, 20 μm. (B) Quantification with ImageJ of donor H2 Ag spots per FDC network on spleens of BALB/c mice transplanted with B6 hearts. (C) Donor H2 Ag spots per FDC network on BALB/c LNs draining B6 skin allografts assessed using ImageJ. (D) Deep SIM of donor H2 Ag spots in FDCs of a BALB/c LN draining a B6 skin graft. Scale bar, 2 μm. ROI, region of interest. (E and F) 2P-microscopy of FDCs in the spleen of a BALB/c mouse transplanted with a B6 heart releasing RFP-sEVs (E) and a BALB/c LN draining a B6 skin graft releasing RFP-sEVs (F). Arrows: graft-RFP-sEVs on the surface (red) or inside (yellow) of FDCs (green). CellTrace Violet-B cells indicate B cell follicles. FDCs were labeled with AF488-CD21/CD35 Ab. Original magnification ×25. Scale bars, 20 μm. (G) Colocalization by confocal microscopy of donor (B6, IA b + H2K b ) allo-Ags in FDCs of LNs draining CMV Cre/+ LSL -RFP-CD63 B6 skin grafted in BALB/c mice. Scale bar, 10 μm. Mander’s colocalization coefficient of graft-sEVs and donor IA b + H2K b Ab analyzed using Imaris. Each triangle represents one FDC network. In (E) and (F), RFP spots per FDC network z stacks were quantified within 90-μm-thick z stacks using Imaris. Experiments were done on 4 spleens per POD (A and B), 6 graft-dLNs per POD (C), 3 graft-dLNs (D), 4 spleens (E), 4 graft-dLNs (F), and 12 graft-dLNs (G). In (B), (C), (E), and (F), comparisons were made by multiple unpaired two-tailed Student’s test. Error bars, means ± SD; * p < 0.05, ** p < 0.01, *** p < 0.001, and **** p < 0.0001; NS, not significant.

    Journal: Cell reports

    Article Title: Transplants foster B cell alloimmunity by relaying extracellular vesicles to follicular dendritic cells

    doi: 10.1016/j.celrep.2025.115832

    Figure Lengend Snippet: (A) Donor H2 Ag in splenic FDCs (inset, arrows) after transplantation of B6 (H2 b ) hearts in BALB/c mice. Original magnification ×200. Scale bars, 20 μm. (B) Quantification with ImageJ of donor H2 Ag spots per FDC network on spleens of BALB/c mice transplanted with B6 hearts. (C) Donor H2 Ag spots per FDC network on BALB/c LNs draining B6 skin allografts assessed using ImageJ. (D) Deep SIM of donor H2 Ag spots in FDCs of a BALB/c LN draining a B6 skin graft. Scale bar, 2 μm. ROI, region of interest. (E and F) 2P-microscopy of FDCs in the spleen of a BALB/c mouse transplanted with a B6 heart releasing RFP-sEVs (E) and a BALB/c LN draining a B6 skin graft releasing RFP-sEVs (F). Arrows: graft-RFP-sEVs on the surface (red) or inside (yellow) of FDCs (green). CellTrace Violet-B cells indicate B cell follicles. FDCs were labeled with AF488-CD21/CD35 Ab. Original magnification ×25. Scale bars, 20 μm. (G) Colocalization by confocal microscopy of donor (B6, IA b + H2K b ) allo-Ags in FDCs of LNs draining CMV Cre/+ LSL -RFP-CD63 B6 skin grafted in BALB/c mice. Scale bar, 10 μm. Mander’s colocalization coefficient of graft-sEVs and donor IA b + H2K b Ab analyzed using Imaris. Each triangle represents one FDC network. In (E) and (F), RFP spots per FDC network z stacks were quantified within 90-μm-thick z stacks using Imaris. Experiments were done on 4 spleens per POD (A and B), 6 graft-dLNs per POD (C), 3 graft-dLNs (D), 4 spleens (E), 4 graft-dLNs (F), and 12 graft-dLNs (G). In (B), (C), (E), and (F), comparisons were made by multiple unpaired two-tailed Student’s test. Error bars, means ± SD; * p < 0.05, ** p < 0.01, *** p < 0.001, and **** p < 0.0001; NS, not significant.

    Article Snippet: CellTrace TM Violet Cell Proliferation Kit, for flow cytometry , Thermo Fisher , Cat # C34557.

    Techniques: Transplantation Assay, Microscopy, Labeling, Confocal Microscopy, Two Tailed Test

    Selective expansion and differentiation of T cells by AP-EVs-Th1 and AP-EVs-Th2 in vitro . (a) Lymph node T cells from C57BL/6 and OT-II transgenic mice were combined in a 1:1 ratio. The combined cells were labeled with CellTrace Violet (CTV) and treated with control EVs or AP-EVs-Th1. (b) The combined cells were co-cultured with AP-EVs-Th1, control EVs, or anti-mouse CD3/CD28 conjugated beads for three days, and proliferation and differentiation of OT-II T cells were analyzed via flow cytometry. (c) Flow cytometric plots of CTV-labeled OT-II (CD45.2/CD45.1) and polyclonal (CD45.1) T cells cultured with anti-mouse CD3/CD28 conjugated beads, AP-EVs-Th1, or control EVs. (d) Percentage of T-bet-expressing CD4 + T cells. (e) Bar graph showing the percentage of T-bet-expressing wild-type (WT) CD4 + T or OT-II T cells under different concentrations of control EVs (black) and AP-EVs-Th1 (orange). (f) The combined cells were co-cultured with AP-EVs-Th2 or control EVs for three days, and the proliferation and differentiation of OT-II T cells were analyzed via flow cytometry. (g) Flow cytometric plots of CTV-labeled OT-II (CD45.2) and polyclonal (CD45.1) T cells cultured with anti-mouse CD3/CD28 conjugated beads, AP-EVs-Th2, or control EVs. (h) Percentage of GATA-3-expressing CD4 + T cells. (i) Bar graph showing the percentage of GATA-3-expressing WT CD4 + or OT-II T cells under different concentrations of control EVs (black) and AP-EVs-Th2 (orange).

    Journal: Drug Delivery

    Article Title: Selective expansion and differentiation of antigen-specific CD4 + T-helper cells by engineered extracellular vesicles

    doi: 10.1080/10717544.2025.2509969

    Figure Lengend Snippet: Selective expansion and differentiation of T cells by AP-EVs-Th1 and AP-EVs-Th2 in vitro . (a) Lymph node T cells from C57BL/6 and OT-II transgenic mice were combined in a 1:1 ratio. The combined cells were labeled with CellTrace Violet (CTV) and treated with control EVs or AP-EVs-Th1. (b) The combined cells were co-cultured with AP-EVs-Th1, control EVs, or anti-mouse CD3/CD28 conjugated beads for three days, and proliferation and differentiation of OT-II T cells were analyzed via flow cytometry. (c) Flow cytometric plots of CTV-labeled OT-II (CD45.2/CD45.1) and polyclonal (CD45.1) T cells cultured with anti-mouse CD3/CD28 conjugated beads, AP-EVs-Th1, or control EVs. (d) Percentage of T-bet-expressing CD4 + T cells. (e) Bar graph showing the percentage of T-bet-expressing wild-type (WT) CD4 + T or OT-II T cells under different concentrations of control EVs (black) and AP-EVs-Th1 (orange). (f) The combined cells were co-cultured with AP-EVs-Th2 or control EVs for three days, and the proliferation and differentiation of OT-II T cells were analyzed via flow cytometry. (g) Flow cytometric plots of CTV-labeled OT-II (CD45.2) and polyclonal (CD45.1) T cells cultured with anti-mouse CD3/CD28 conjugated beads, AP-EVs-Th2, or control EVs. (h) Percentage of GATA-3-expressing CD4 + T cells. (i) Bar graph showing the percentage of GATA-3-expressing WT CD4 + or OT-II T cells under different concentrations of control EVs (black) and AP-EVs-Th2 (orange).

    Article Snippet: Lymph node T cells isolated from C57BL/6 and OT-II transgenic mice were stained with 1 μM CellTrace Violet (CTV) Cell Proliferation Kit (Thermo Fisher Scientific, Waltham, MA) for flow cytometry and incubated at 37 °C for 3 min.

    Techniques: In Vitro, Transgenic Assay, Labeling, Control, Cell Culture, Flow Cytometry, Expressing