fitc anti17 mouse cd80  (Multi Sciences (Lianke) Biotech Co Ltd)

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

Journal: Journal of Extracellular Vesicles

Article Title: High‐Yield Outer Membrane Vesicles Derived From Probiotics as a Nanoplatform for Precise Treatment and Prophylaxis of Pseudomonas aeruginosa Infection

doi: 10.1002/jev2.70194

Figure Lengend Snippet: In vitro immune response of antigen‐presenting cells activated by FI‐mOMVs. (a) Confocal microscopy images of antigen uptake by BMDCs after incubation with the indicated formulations for 6 h. The cell immunofluorescence staining was performed by the marker protein CD11c on the cell membrane. The cell nuclei were stained with DAPI. Scale bar, 100 µm. (b, c) Flow cytometry was used to detect the uptake efficiency of FI antigen after incubation with BMDCs (b), and the uptake level was quantified (c) ( n = 3). (d) The uptake efficiency of BMDCs on unbroken FI‐mOMVs and broken FI‐mOMVs was detected by flow cytometry ( n = 3). (e, f) Flow cytometry was performed to measure the percentage of CD80 + (e) or CD86 + (f) cells in CD11c + BMDCs after incubation with the indicated formulations for 24 h ( n = 3). (g) FITC‐FI, DiI‐mOMVs and DiI‐FI‐mOMVs were incubated with PMs for 6 h. The cell immunofluorescence staining was performed by the marker protein F4/80 on the cell membrane. After DAPI staining, laser scanning confocal microscopy was used to detect the uptake of FI‐mOMVs by PMs. Scale bar, 100 µm. (h, i) Flow cytometry was used to quantitatively detect the uptake level of FI antigen by PMs (h), and quantitative analysis was performed (i) ( n = 3). (j) The activities of immune‐related enzymes ACP, LDH, iNOS in PMs were detected by ELISA after different treatments for 12 h ( n = 3). (k) Production of TNF‐α, IL‐6, TGF‐β and IL‐10 in the cell supernatant measured by ELISA after different treatments for 12 h ( n = 3). The data are presented as mean ± SEM. Statistical analysis was performed by a two‐tailed unpaired Student's t ‐test, and differences were considered significant at p < 0.05. ns, not significant; * p < 0.05; ** p < 0.01; *** p < 0.001 and **** p < 0.0001.

Article Snippet: The BMDCs were collected and then labelled by PE anti‐mouse CD11c (cat# PE‐65130, Proteintech, USA), FITC anti‐mouse CD80 (cat# FITC‐65076, Proteintech, USA) and FITC anti‐mouse CD86 (cat# FITC‐65068, Proteintech, USA).

Techniques: In Vitro, Confocal Microscopy, Incubation, Immunofluorescence, Staining, Marker, Membrane, Flow Cytometry, Enzyme-linked Immunosorbent Assay, Two Tailed Test