Journal: bioRxiv

Article Title: Adjuvant Discovery via a High Throughput Screen using Human Primary Mononuclear Cells

doi: 10.1101/2022.06.17.496630

Figure Lengend Snippet: Key resource table

Article Snippet: CD86 Antibody, anti-human, PE, REAfinityTM (Clone- REA968) , Miltenyi Biotec , Order Number: 130-116-160.

Techniques: Recombinant, Sterility, Injection, Modification, Staining, Saline, Enzyme-linked Immunosorbent Assay, Multiplex Assay, Software

Journal: Central European Journal of Immunology

Article Title: Macrophage M2 polarization induced by ANKRD22 in lung adenocarcinoma facilitates tumor angiogenesis

doi: 10.5114/ceji.2025.149372

Figure Lengend Snippet: Fig. 2. Cont. F-H) Flow cytometry was used to detect the effects of ANKRD22 overexpression or knockdown on the proportions of CD68+CD86+ and CD68+CD206+ macrophages. All experiments were conducted with 3 independent replicates. The data are presented as mean ± standard deviation. *p < 0.05

Article Snippet: The cells were incubated with PerCP Anti-Human CD68 Antibody (1 : 100; 333813, BioLegend, USA), PE Anti-Human CD86 Antibody (1 : 100; E-AB-F1012D, Elabscience, China), and/or FITC AntiHuman CD206/MMR Antibody (1 : 100; E-AB-F1161C, Elabscience, China) for 15 min. After washing with PBS, the cells were resuspended, and the flow cytometry data were collected and analyzed using Agilent’s flow analysis software.

Techniques: Flow Cytometry, Over Expression, Knockdown, Standard Deviation

Journal: Central European Journal of Immunology

Article Title: Macrophage M2 polarization induced by ANKRD22 in lung adenocarcinoma facilitates tumor angiogenesis

doi: 10.5114/ceji.2025.149372

Figure Lengend Snippet: Fig. 2. Cont. I) Flow cytometry was used to detect the ef- fects of ANKRD22 overexpression or knockdown on the pro- portions of CD68+CD86+ and CD68+CD206+ macrophages. J) WB was used to detect the effects of ANKRD22 over- expression or knockdown on the protein expression levels of CD80 and CD163. All experiments were conducted with 3 independent replicates. The data are presented as mean ± standard deviation. *p < 0.05

Article Snippet: The cells were incubated with PerCP Anti-Human CD68 Antibody (1 : 100; 333813, BioLegend, USA), PE Anti-Human CD86 Antibody (1 : 100; E-AB-F1012D, Elabscience, China), and/or FITC AntiHuman CD206/MMR Antibody (1 : 100; E-AB-F1161C, Elabscience, China) for 15 min. After washing with PBS, the cells were resuspended, and the flow cytometry data were collected and analyzed using Agilent’s flow analysis software.

Techniques: Flow Cytometry, Over Expression, Knockdown, Expressing, Standard Deviation

Journal: Journal for Immunotherapy of Cancer

Article Title: SP140 inhibits STAT1 signaling, induces IFN-γ in tumor-associated macrophages, and is a predictive biomarker of immunotherapy response

doi: 10.1136/jitc-2022-005088

Figure Lengend Snippet: Overexpression of SP140 induces the production of inflammatory cytokines and chemokines and reprogram macrophages to induce TAM-mediated tumor cytotoxicity. (A, B) Control CRISPR/dCAS9 (control) or SP140 CRISPR/dCAS9 (SP140 overexpression) was transfected in naïve macrophages, and the expression of CD80 and CD86 was quantified by flow cytometry. (C–E) SP140 CRISPR/dCAS9 (SP140 OE) or scramble control (control CRISPR/dCAS9) were introduced to the naïve macrophages, and a multiplex fluorescence BioLegend assay was used to identify the levels of CXCL10, IFN-γ, and IL-12 (P70) in the supernatant after 48 hours. LPS (100 nM) was administered after 24 hours. (F–I) Correlation of SP140 expression and IFNG, CXCL10, IL-12B, and IL-12A levels in over 500 HNSCCs in the TCGA dataset. (J) Control CRISPR/dCAS9 or SP140 CRISPR/dCAS9 was introduced in TAMs isolated from syngeneic HNSCC tumor and cocultured with murine SCC7 cells. Cell viability after 48 hours of coculture was quantified with a quantitative viability assay kit. Data are presented based on the fold change of non-treated control. *P<0.05, ***P<0.001, ****P<0.0001. HNSCC, head and neck squamous cell carcinoma; IFN-γ, interferon gamma; IL, interleukin; SP140, speckled protein 140; TAM, tumor-associated macrophage; TCGA, The Cancer Genome Atlas.

Article Snippet: A flow cytometry panel consisting of human CD80 (PE, BioLegend), human CD86 (BV711, BioLegend), and human CD206 (PerCP-Cy5.5, BioLegend) was used for the characterization of macrophages.

Techniques: Over Expression, Control, CRISPR, Transfection, Expressing, Flow Cytometry, Multiplex Assay, Fluorescence, Isolation, Viability Assay