anti human cd206 15 2  (Proteintech)

Journal: Scientific Reports

Article Title: CCL8 suppresses ovarian cancer progression via M1 macrophage polarization and NF-κB-mediated apoptosis

doi: 10.1038/s41598-025-32883-1

Figure Lengend Snippet: CCL8 promotes THP1 polarization to M1-type macrophages and inhibits OC cell growth and metastasis. ( A ) THP1-derived macrophages were stimulated with CCL8 for 0 h, 24 h, and 48 h, after which the mRNA expression of M1 macrophage-associated markers (IL-1β, IL-6, CXCL10) and M2 macrophage-associated markers (CD206, CD163, ARG, IL-10) was examined.GAPDH was used as a loading control ( n = 3 independent replicates; two-way ANOVA). ( B ) CD80 and CD206 protein expression in THP1 after stimulation with CCL8, and the quantitative analysis by ImageJ-win64. GAPDH was used as a loading control( n = 3 independent replicates; two-way ANOVA). ( C ) Flow cytometric analysis of CD86-positive and CD206-positive cells in THP1 cells following CCL8 treatment. ( D ) Supernatants of THP1 cells treated with PMA and then stimulated with CCL8 (50 ng/mL) or vehicle (PBS) for 48 h were collected and used to treat ES2 cells for 48 h. ( E ) Colony formation analysis of ES2 cells treated with NC-CM or CCL8-CM ( n = 3 independent replicates; Student’s t-test). ( F ) CCK-8 analysis of growth rates of ES2 cells treated with NC-CM or CCL8-CM ( n = 3 independent replicates; Student’s t-test). ( G , H ) Transwell migration ( G ) and invasion ( H ) analysis of ES2 cells treated with NC-CM or CCL8-CM ( n = 3 independent replicates; Student’s t-test). ( I ) Representative Western blot gel files and pooled data showing the proteins expression of EMT, stemness and proliferation indicator in ES2 cells treated with NC-CM or CCL8-CM, and the quantitative analysis by ImageJ-win64. GAPDH was used as a loading control ( n = 3 independent replicates; two-way ANOVA). Scale bar:100 μm.* P <.05, ** P <.01, **** P <.0001.

Article Snippet: 1 × 106 THP1 cells were surface co-stained with 5 μL FITC Anti-Human CD206 (Anti-Human CD206 Rabbit Recombinant Antibody, 98458-3-RR, Proteintech, China) and 5 μL PE Anti-Human CD86 (BU63) (PE-65165, Proteintech, China) or left unstained, then incubated at 4 °C in the dark for 40 min. Wash cells with 1 × PBS, centrifuge at 500 × g for 5 min, and discard the supernatant.

Techniques: Derivative Assay, Expressing, Control, CCK-8 Assay, Migration, Western Blot