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: Discover Oncology

Article Title: Integrating multi-omics data to optimize immunotherapy in endometrial cancer: a comprehensive study

doi: 10.1007/s12672-025-02978-2

Figure Lengend Snippet: Effect of TAMs phenotype on immunologic efficacy in EC patients and proliferation and migration for Ishikawa cells. A Immunohistochemistry (IHC) staining of M1-like TAMs (CD86 + ) and M2-like TAMs (CD206 + ) in tumor tissues from EC patients. The bar graphs on the right show the IHC staining scores (*** P ≤ 0.001). B Flow cytometry analysis of M1-like TAMs (CD86 + ) and M2-like TAMs (CD206 + ) in PBMCs. The left panel shows representative flow plots, and the bar graphs on the right quantify the percentages (*** P ≤ 0.001). C Experimental design for TAMs polarization and co-culture system. D CCK-8 assay measuring cell viability of Ishikawa cells co-cultured with M1 and M2-like TAMs for 24, 48 and 72 h. The bar graphs show the percentage of viable cells (* P < 0.05, ** P ≤ 0.01). E Transwell migration assay of Ishikawa cells co-cultured with M1 and M2-like TAMs. The bar graphs on the right quantify the number of migrated cells (* P < 0.05, ** P ≤ 0.01)

Article Snippet: The cells were then stained with 4 μL of CD206 antibody (FITC-65155, Proteintech, Hunan, China) and 5 μL of CD86 antibody (PE-65165, Proteintech, Hunan, China), followed by 40 min of incubation in the dark at 4 °C.

Techniques: Migration, Immunohistochemistry, Flow Cytometry, Co-Culture Assay, CCK-8 Assay, Cell Culture, Transwell Migration Assay

Journal: Redox Biology

Article Title: Regulation of type I interferon responses by mitochondria-derived reactive oxygen species in plasmacytoid dendritic cells

doi: 10.1016/j.redox.2017.07.016

Figure Lengend Snippet: Phenotypical analysis of AMA-exposed GEN2.2 cell. Cells were treated with AMA (0.5 μg/ml) and CpG-A (1 μM) separately and in combination or were left untreated. Following 6 h of stimulation the expression levels of CD40 (A), CD80 (B), CD86 (C) and HLA-DQ (D) cell surface proteins were analyzed by flow cytometry. Relative fluorescence intensity values were calculated using the respective isotype-matched control antibodies. The bars represent fold changes compared to the untreated control and data are expressed as the mean ± SD of four independent experiments. *p < 0.05 vs. control. AMA: Antimycin-A.

Article Snippet: After 24 h of treatments the phenotypical analysis of the cells was performed by flow cytometry using FITC-labeled monoclonal antibodies against CD40 and CD80 (Cat. no. 334306 and 305206 both from BioLegend, San Diego, CA, USA), PE-labeled CD86 (Cat. no. FAB141P from R&D System, Minneapolis, MN, USA), HLA-DQ (Cat. no. 318106 from BioLegend) and isotype-matched control antibodies (all from BioLegend).

Techniques: Expressing, Flow Cytometry, Fluorescence