Journal: NPJ Vaccines

Article Title: A KIF20A-based thermosensitive hydrogel vaccine effectively potentiates immune checkpoint blockade therapy for hepatocellular carcinoma

doi: 10.1038/s41541-024-01060-2

Figure Lengend Snippet: a CD80 and CD86 levels of BMDCs in flow cytometry (The gating strategy was shown in the Supplementary Fig. ). b Quantification of the proportion of CD80 + CD86 + BMDCs. c MHC-I level of BMDCs indicated by flow cytometry (The gating strategy was shown in the Supplementary Fig. ). d Quantification of the proportion of MHC-I + BMDCs. e MHC-II level of BMDCs in flow cytometry (The gating strategy was shown in the Supplementary Fig. ). f Quantification of MHC-II + BMDCs. g CFDA-SE level in CD3 + T cells (The gating strategy was shown in the Supplementary Fig. ). h Quantification of T cell proliferation rate. i CLSM images of infiltrated CD8a + T cells into Hepa1-6 cell spheroids. j Quantification of T cell infiltration.

Article Snippet: Cells were subsequently stained with APC-conjugated anti-mouse CD11c, FITC-conjugated anti-mouse CD80, and PE-conjugated anti-mouse CD86 antibodies (Elabscience, China) for flow cytometry analysis (Beckman, USA).

Techniques: Flow Cytometry

Journal: NPJ Vaccines

Article Title: A KIF20A-based thermosensitive hydrogel vaccine effectively potentiates immune checkpoint blockade therapy for hepatocellular carcinoma

doi: 10.1038/s41541-024-01060-2

Figure Lengend Snippet: a Scheme illustration of in vivo treatment and monitoring (Created by the authors with Photoshop v22.5.6). b Bioluminescence images of tumor-bearing mice in different groups. c Relative signal intensity of tumors in different groups. d Body weight of the mice in different groups. e Flow cytometry analyzing CD80 and CD86 in CD11c + dendritic cells in mouse inguinal lymph nodes (Gating strategy was shown in the Supplementary Fig. ). f Quantification of the percentage of CD80 + CD86 + cells in CD11c + dendritic cells. g Flow cytometry analyzing CD8a and CD4 of T cells (CD45 + CD3 + ) in tumor tissues (Gating strategy was shown in the Supplementary Fig. ). h Quantification of the percentage of CD8a + cells in T cells.

Article Snippet: Cells were subsequently stained with APC-conjugated anti-mouse CD11c, FITC-conjugated anti-mouse CD80, and PE-conjugated anti-mouse CD86 antibodies (Elabscience, China) for flow cytometry analysis (Beckman, USA).

Techniques: In Vivo, Flow Cytometry

Journal: STAR Protocols

Article Title: A protocol for high-throughput screening for immunomodulatory compounds using human primary cells

doi: 10.1016/j.xpro.2023.102405

Figure Lengend Snippet: Representative schematic of the high throughput flow cytometry system A total of 5 μL of antibodies diluted 1:200 in PBS were added to each well containing 30 μL of cells (50,00 cells/well). This flow cytometry system captures the total events or cells per well in addition to the antibody activity, which measures markers including the surface receptors Ox40, CD80/86, and HLA-DR. The flow cytometry approach also identifies B cells, T cells, and monocytes.

Article Snippet: CD80 Antibody, anti-human, PE, REAfinityTM (Clone- REA661). Diluted 1:200 with dPBS by volume. , Miltenyi Biotec , Order Number: 130-123-253.

Techniques: High Throughput Screening Assay, Flow Cytometry, Activity Assay

Journal: STAR Protocols

Article Title: A protocol for high-throughput screening for immunomodulatory compounds using human primary cells

doi: 10.1016/j.xpro.2023.102405

Figure Lengend Snippet:

Article Snippet: CD80 Antibody, anti-human, PE, REAfinityTM (Clone- REA661). Diluted 1:200 with dPBS by volume. , Miltenyi Biotec , Order Number: 130-123-253.

Techniques: Recombinant, Sterility, Modification, Staining, Saline, Multiplex Assay, Software

Journal: Journal of Inflammation (London, England)

Article Title: Tissue-plasminogen activator effects on the phenotype of splenic myeloid cells in acute inflammation

doi: 10.1186/s12950-024-00375-0

Figure Lengend Snippet: References of flow cytometry antibodies used for immunophenotyping

Article Snippet: CD80 , REA983 , APC-Vio770 , 130–116-463 , human IgG1/REA293 , Miltenyi Biotec.

Techniques: Flow Cytometry, Clone Assay

Journal: Journal of Inflammation (London, England)

Article Title: Tissue-plasminogen activator effects on the phenotype of splenic myeloid cells in acute inflammation

doi: 10.1186/s12950-024-00375-0

Figure Lengend Snippet: Effects of LPS treatment on the phenotype of splenic macrophages from tPA −/− mice. A Representative flow cytometry gating strategy used for quantification of spleen macrophages (F4/80 + CD11b + ), expressing MHCII molecules or not (F4/80 + CD11b + MHCII +or− ) and costimulatory molecules (F4/80 + CD11b + MHCII +or− CD80 + CD86 + ). B Frequency of macrophages (Sham WT n = 9; Sham tPA −/− ; LPS WT; LPS tPA −/− n = 10). C Frequency of MHCII − macrophages (F4/80 + CD11b + MHCII − ) and of costimulatory molecule expressing cells (F4/80 + CD11b + MHCII − CD80 + CD86 + ), (Sham WT n = 9; Sham tPA −/− ; LPS WT; LPS tPA −/− n = 10). D Frequency of MHCII + macrophages (F4/80 + CD11b + MHCII + ), (Sham WT n = 9; Sham tPA −/− ; LPS WT; LPS tPA −/− n = 10), MFI quantification of MHCII on MHCII + macrophages (Sham WT n = 9; Sham tPA −/− ; LPS WT; LPS tPA −/− n = 10) and frequency of MHCII + macrophages expressing costimulatory molecules (F4/80 + CD11b + MHCII + CD80 + CD86 + ), (Sham WT n = 9; Sham tPA −/− ; LPS WT; LPS tPA −/− n = 10). E MFI quantification of CD80 (Sham WT n = 8; Sham tPA −/− ; LPS WT n = 10; LPS tPA −/− n = 9), CD86 (Sham WT n = 8; Sham tPA −/− n = 9, LPS WT; LPS tPA −/− n = 10) and CD11b molecules (Sham WT n = 8; Sham tPA −/− ; LPS WT; LPS tPA −/− n = 10) on MHCII − macrophages. F MFI quantification of CD80 (Sham WT n = 8; Sham tPA −/− ; LPS WT n = 10; LPS tPA −/− n = 9), CD86 and CD11b molecules (Sham WT n = 9; Sham tPA −/− ; LPS WT; LPS tPA −/− n = 10) on MHCII + macrophages. Data are shown as individual animals with mean ± SD, two-way ANOVA with Bonferroni’s post-hoc

Article Snippet: CD80 , REA983 , APC-Vio770 , 130–116-463 , human IgG1/REA293 , Miltenyi Biotec.

Techniques: Flow Cytometry, Expressing

Journal: Journal of Inflammation (London, England)

Article Title: Tissue-plasminogen activator effects on the phenotype of splenic myeloid cells in acute inflammation

doi: 10.1186/s12950-024-00375-0

Figure Lengend Snippet: cDC phenotype was modulated in an inflammatory setting. A Representative flow cytometry gating strategy used for quantification of total DCs (F4/80 − CD11c + ) and cDCs: cDC1 (F4/80 − CD11c + CD11b − MHCII + ) and cDC2 (F4/80 − CD11c + CD11b + MHCII + ). B Quantification of total DC frequency (Sham WT n = 8; Sham tPA −/− ; LPS WT; LPS tPA −/− n = 10). C Frequency of cDC1, MFI of MHCII molecules on cDC1 and frequency of CD80 + CD86 + cDC1 (Sham WT n = 9; Sham tPA −/− ; LPS WT; LPS tPA −/− n = 10). D Frequency of cDC2 and CD80 + CD86 + cDC2, MFI of MHCII and CD11b on cDC2 (Sham WT n = 9; Sham tPA −/− ; LPS WT; LPS tPA −/− n = 10). Data are shown as individual animals with mean ± SD, two-way ANOVA with Bonferroni’s post-hoc

Article Snippet: CD80 , REA983 , APC-Vio770 , 130–116-463 , human IgG1/REA293 , Miltenyi Biotec.

Techniques: Flow Cytometry