Multiparameter Flow Cytometry Analysis

Single cell suspensions of spleens were blocked with anti-mouse CD16/32 (BioLegend) and surface stained with the indicated markers, and evaluated by flow cytometric analysis as described^{18 (link),29 (link),35 (link)–37 (link)}. The following Abs were used; anti-CD45 (clone 30-F11, Invitrogen), anti-PD-1 (clone 29F.1A12, BioLegend), anti-CX3CR1 (clone SA011F11, BioLegend), anti-KLRG1 (clone 2F1/KLRG1, BioLegend), anti-CD4 (clone GK1.5, BioLegend), anti-CD8α (clone 53-6.7, BD Biosciences), and anti-CD3 (clone 145-2C11, BioLegend). LIVE/DEAD Fixable Near-IR Dead Cell Stain kit (Thermo Fisher Scientific)-stained cells were excluded from the analysis. Samples were acquired by Fortessa (BD Biosciences) cytometers, and analyzed with FlowJo software (Treestar).

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Yokoi T., Oba T., Kajihara R., Abrams S.I, & Ito F. (2021). Local, multimodal intralesional therapy renders distant brain metastases susceptible to PD-L1 blockade in a preclinical model of triple-negative breast cancer. Scientific Reports, 11, 21992.

Publication 2021

anti-mouse CD16/32 anti-CD45 (clone 30-F11, anti-PD-1 (clone 29F.1A12, anti-CX3CR1 (clone SA011F11, anti-KLRG1 (clone 2F1/KLRG1, anti-CD4 (clone GK1.5, anti-CD8α (clone 53-6.7, anti-CD3 (clone 145-2C11, LIVE/DEAD Fixable Near-IR Dead Cell Stain kit Fortessa

Anti cd3 Cells Clone Flow cytometric Klrg1 Mouse

Corresponding Organization :

Other organizations : Roswell Park Comprehensive Cancer Center

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Variable analysis

independent variables

Blocking with anti-mouse CD16/32 (BioLegend)
Surface staining with the indicated markers

dependent variables

Flow cytometric analysis of cells

control variables

LIVE/DEAD Fixable Near-IR Dead Cell Stain kit (Thermo Fisher Scientific)-stained cells were excluded from the analysis

controls

Positive control: Not explicitly mentioned
Negative control: Not explicitly mentioned

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