celltrace violet (ctv) cell proliferation kit  (Thermo Fisher)

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

Journal: bioRxiv

Article Title: LAG3 marks activated but hyporesponsive NK cells

doi: 10.1101/2024.12.18.629184

Figure Lengend Snippet: (A) C57BL/6 mice (n=4 for each day) were injected intraperitoneally with 200ug of Poly (I:C) and sacrificed in kinetic. The percentage of Sca1+, LAG3+ and CD69+ splenic NK cells was analyzed by flow cytometry. (B) The percentage of LAG3+ among Sca1+ and Sca1-NK cells on Day2 was analyzed by flow cytometry (n=5). (C-G) C57BL/6 mice (n=2 for each experiment) were injected intraperitoneally with 200ug of Poly (I:C) and sacrificed on Day2. Splenic NK cells were stained for the phospho-epitope pS6 Ser235/236 (C) and for pAkt S473 (D). Overlays of representative histograms are shown (left panels). The MFI values are indicated on the graph and represented on the right panel for each individual, n=6, 3 experiments in total. (E) NK cells were stained with Mitotracker Green and CellRox. The MFI values for each mouse (n=7, 3 experiments in total) for the analyzed marker are represented. (F) The MFI of CD98 (left) or CD71 (right) was determined by flow cytometry on LAG3+ and LAG3-NK cells (n=3). Due to the small sample size, we did not calculate p-values. (G) SSC-A and FSC-A parameters on NK cells were measured by flow cytometry in LAG3- and LAG3+ NK cells (n=5). (H) Splenic NK cells from C57BL/6 mice (n=6, 3 experiments) were isolated, stained with Cell Trace Violet (CTV) and cultured with 100ng/mL of IL-15 for 3 days. The MFI of CTV was analyzed by flow cytometry and the overlays of representative histograms are shown (left panel). The percentage of divided NK cells for each individual was calculated using the proliferation modeling tool on the FlowJo software. All the data were analyzed using a paired non-parametric Wilcoxon test. p-values are indicated on each graph.

Article Snippet: Isolated NK cells were stained with 0.6µM Invitrogen CellTrace TM Violet (CTV) Cell Proliferation Kit according to manufacturer’s instructions.

Techniques: Injection, Flow Cytometry, Staining, Marker, Isolation, Cell Culture, Software

Journal: iScience

Article Title: Combined TLR2/TLR4 activation equip non-mucosal dendritic cells to prime Th1 cells with gut tropism

doi: 10.1016/j.isci.2024.111232

Figure Lengend Snippet:

Article Snippet: The cells from the cultures with DCs stimulated with MPLA alone were stained with CFSE (CellTrace CFSE Cell Proliferation Kit, for flow cytometry; Invitrogen, #C34554) and the cells from MPLA+P3C activated cultures were stained with CTV (CellTrace Violet Cell Proliferation Kit, for flow cytometry; Invitrogen, #C34557).

Techniques: Recombinant, Lysis, Cell Isolation, Staining, Enzyme-linked Immunosorbent Assay, Software