cd80 fitc  (Miltenyi Biotec)

Journal: Cellular and Molecular Life Sciences: CMLS

Article Title: Cathelicidin-Ka, the first frog-derived TLR2 and TLR4 agonist, induces macrophage activation and promotes inflammation

doi: 10.1007/s00018-025-06068-y

Figure Lengend Snippet: Cath-Ka promotes macrophage activation and M1 polarization. ( A ) Representative morphology images of RAW264.7 cells. RAW264.7 cells were treated with PBS, Cath-Ka (2.5–20 µM), or LPS (100 ng/mL) for 24 h before images were captured. Scale bar = 50 μm. ( B ) Viability of RAW264.7 cells incubated with PBS, Cath-Ka (1.25–20 µM), or LPS (100 ng/mL) for 24 h. ( C , D ) RT-qPCR analysis of pro-inflammatory cytokines mRNA expression in RAW264.7 cells. Cells were incubated with PBS or Cath-Ka at the indicated concentrations for 2 h ( C ) or 4 h ( D ) before RT-qPCR analysis. ( E ) ELISA analysis of pro-inflammatory cytokine secretion. RAW264.7 cells were incubated with PBS, Cath-Ka (1.25–20 µM), or LPS (100 ng/mL) for 24 h before ELISA assays. ( F ) Effect of PMB (10 µg/mL) on TNF-α production in RAW264.7 cells stimulated by Cath-Ka (2.5–10 µM) or LPS (100 ng/mL). ( G ) Representative flow cytometry plots (left) and statistical analysis (right) of intracellular ROS. RAW264.7 cells were incubated with PBS, Cath-Ka (1.25–20 µM), or LPS (100 ng/mL) for 24 h before flow cytometry analysis. ( H ) Representative flow cytometry histograms and ( I ) statistical analysis of CD80, CD86, MHC II, and CD206 expression. RAW264.7 cells were incubated with PBS, Cath-Ka (1.25–20 µM), or LPS (100 ng/mL) for 24 h before their CD80, CD86, MHC II and CD206 expression was measured by flow cytometry. Data are expressed as mean ± SD ( n = 3). * P < 0.05, ** P < 0.01, *** P < 0.001; ns, not significant

Article Snippet: PE-anti-mouse MHC-II (#F21IIAE02) and FITC-anti-mouse CD80 (#F2108001) were from MultiSciences.

Techniques: Activation Assay, Incubation, Quantitative RT-PCR, Expressing, Enzyme-linked Immunosorbent Assay, Flow Cytometry