violet 450 anti mouse cd4 antibody  (Elabscience Biotechnology)

Journal: Journal of Virology

Article Title: A novel nanoparticle vaccine, based on S1-CTD, elicits robust protective immune responses against porcine deltacoronavirus

doi: 10.1128/jvi.00674-25

Figure Lengend Snippet: CTDnps vaccine elicited robust antibody and T cell responses in mice. ( A ) Schematic diagram of the mice immunization procedure. Five mice per group received prime/boost vaccination with various vaccines at Weeks 0 and 4. Serum samples were collected every 2 weeks. ( B ) Titers of antigen-specific IgG antibodies were measured in serum samples by iELISA. ( C ) Titers of NAbs were measured in serum samples from mice at Week 8 by VNT. ( D ) Titers of antigen-specific IgG1 and IgG2a for each immunization group. Serum samples from mice at Week 8 were tested. IL-4 ( E ) and IFN-γ ( F ) concentrations were measured by double-antibody sandwich ELISA. ( G, H ) The percentage of CD3 + CD4 + and CD3 + CD8 + T lymphocytes among spleen lymphocytes was determined by flow cytometry. ( I ) The percentage of CD4 + α4β7 + T lymphocytes among spleen lymphocytes was determined by flow cytometry. ( J, K ) The percentage of CD4 + and CD8 + Tcm among spleen lymphocytes was determined by flow cytometry. Data are mean ± SD. Asterisks indicate significant differences (*, P < 0.05; **, P < 0.01; ***, P < 0.001; and ns, P > 0.05).

Article Snippet: At week 8, spleen lymphocytes of mice were isolated, cell concentration was adjusted to 1 × 10 7 cells/mL, and 10 μL of FITC anti-mouse CD3 antibody, 10 μL of Violet 450 anti-mouse CD4 antibody, and 10 μL of PerCP/Cyanine5.5 anti-mouse CD8a antibody (Elabscience, Wuhan, China) were added to 200 μL of splenocytes for analysis of CD4 + and CD8 + T cells.

Techniques: Vaccines, Sandwich ELISA, Flow Cytometry

Journal: Nature Communications

Article Title: CD276-dependent efferocytosis by tumor-associated macrophages promotes immune evasion in bladder cancer

doi: 10.1038/s41467-024-46735-5

Figure Lengend Snippet: a UMAP plot showing 3 clusters of 2967 macrophages (indicated by colors). b Heatmap of signature genes for macrophages clusters. Each cell cluster is represented by four specifically expressed genes. c KEGG pathway enrichment analysis using the characteristic genes of macrophage subpopulation. d Bar plots of number of the subclusters of macrophages in control and cKO male mice. e Volcano plot displaying the −Log10 P vs Log2 fold-change of genes differentially expressed between control and cKO in macrophages. f KEGG pathway enrichment analysis using the intergroup differential gene of macrophages. g Western blot of CLTC, LIPA, LAPTM5 and LAMP2 in macrophages from control and cKO groups using GAPDH as loading control. h Western blot of CCR1, CCR5, CX3CR1 and CXCR4 in macrophages from control and cKO groups using GAPDH as loading control. i Representative flow cytometry plots (left) and statistical analysis of MHCII + macrophages (right) in control and cKO groups. Data are presented as mean ± SD ( n = 6). P value was calculated by two-tailed unpaired Student’s t test. j Western blot of CXCL9 in MHCII + TAMs and MHCII - TAMs using GAPDH as loading control. k Representative flow cytometry plots (left) and statistical analysis of CD3 + CD4 + T cells and CD3 + CD8 + cells (right) in control and cKO groups. Data are presented as mean ± SD ( n = 6). P value was calculated by two-tailed unpaired Student’s t test. l Representative immunofluorescence (IF) staining images of CD8 (red) and GZMA (green, right). Statistical analysis of the ratio of CD8 + GZMA + cells to CD8 + cells (CD8T killing capacity) in control and cKO male mice (left). Scale bar, 50 μm. Data are expressed as mean ± SD ( n = 6). P values were calculated by two-tailed unpaired Student’s t test.

Article Snippet: Anti-Mouse CD11C PE-CY7 (25-0114-81, ThermoFisher, 1:200); Anti-Mouse F4/80 PCP-CY5.5 (45-4801-80, Thermo Fisher, 1:200); Anti-Mouse Ly-6G (Gr-1) Alexa Fluor® 700 (56-5931-82, ThermoFisher, 1:200); Annexin V-APC/7-AAD Apoptosis kit (AP105, LIANKE, 1:200); Anti-Mouse MHC Class II (I-A/I-E) eFluor® 450 (48-5321-82, ThermoFisher, 1:200); Anti-Mouse CD45 PE-CY7(60-0451, Tonbo, 1:200); Anti-Mouse CD3 APC (20-0032, Tonbo, 1:200); Anti-Mouse CD4 violet Fluor TM 450 (75-0041, Tonbo, 1:200); Anti-Mouse CD8a FITC(35-0081, Tonbo, 1:200) and Ghost Dye TM Red 780 (13-0865, Tonbo, 1:100) were the antibodies and dyes used for flow cytometry (CytoFLEX, Beckman Coulter).

Techniques: Control, Western Blot, Flow Cytometry, Two Tailed Test, Immunofluorescence, Staining