murine recombinant c5a  (R&D Systems)

Journal: Cells

Article Title: Ly6C hi Monocytes Are Metabolically Reprogrammed in the Blood during Inflammatory Stimulation and Require Intact OxPhos for Chemotaxis and Monocyte to Macrophage Differentiation

doi: 10.3390/cells13110916

Figure Lengend Snippet: Reagent and resource list.

Article Snippet: Recombinant murine C5a , PeproTech , 315-40.

Techniques: Generated, Recombinant, Isolation, Reverse Transcription, Expressing, Cytometry, Software

Journal: Cells

Article Title: Ly6C hi Monocytes Are Metabolically Reprogrammed in the Blood during Inflammatory Stimulation and Require Intact OxPhos for Chemotaxis and Monocyte to Macrophage Differentiation

doi: 10.3390/cells13110916

Figure Lengend Snippet: ATP production from oxidative phosphorylation is essential for monocyte and macrophage chemotaxis. ( A ) Schematic of the mammalian electron transport chain. BioGel-elicited myeloid cells were incubated with ( B ) oligomycin (1 µM), ( C ) Bay87-2243 (100 nM), ( D ) 3-nitropropionic (1 mM), ( E ) atovaquine (5 µM), or ( F ) antimycin A (0.5 µM) and a vehicle for 15 min before being added to the upper chamber (4 × 10 5 /well) of a CIM-16 plate and allowed to migrate for 6 h toward murine CCL2 (10 nM). The cell index was measured at 30 s intervals for 6 h and quantified through Max–Min or area under the curve (AUC) analysis. BMDMs were incubated with ( G ) oligomycin (1µM), ( H ) 2-deoxyglucose (50 mM), or ( I ) etomixir (3µM) or a vehicle for 15 min before being added to the upper chamber (1 × 10 5 /well) of a CIM-16 plate and allowed to migrate for 6 h toward murine C5a (10 nM). ( J ) Human monocytes isolated from buffy coats. A total of 4 × 10 5 /well were incubated with oligomycin (1 µM) or a vehicle for 15 min before being added to the upper chamber of a CIM-16 plate and allowed to migrate for 8 h toward human CCL2 (10 nM). Migration was measured with Max–Min analysis and area under the curve (AUC) analysis. Statistical analysis was conducted via one-way ANOVA with Dunnett’s multiple comparison post-test. * p = 0.05, ** p = 0.01.

Article Snippet: Recombinant murine C5a , PeproTech , 315-40.

Techniques: Chemotaxis Assay, Incubation, Isolation, Migration, Comparison

Journal: bioRxiv

Article Title: The membrane-actin linkers ezrin, radixin, and moesin are dispensable for macrophage migration and cortex mechanics

doi: 10.1101/2023.07.27.550674

Figure Lengend Snippet: A. Schematic representation of 2D chemotaxis assay toward C5a. WT and ERM-tKO#1 cells migrating along a C5a gradient in the x axis. See also movie 6. B. Snapshot of WT and ERM-tKO#1 macrophages migrating toward C5a gradient (on the right) with migratory tracks representing cell trajectories during 90min. Tracks are color coded according to their directionality. Scale bar: 50µm. C. Migratory tracks of WT and ERM-tKO#1 macrophages with origins set at (0,0). D. Quantification of the median velocity, the directionality, and the forward migration index in the x axis (FMIx, used as a chemotaxis indicator) of each migratory track. The medians of 3 independent experiments are represented and used for statistical analysis.

Article Snippet: A C5a gradient (R&D systems #2150C5025) was then generated by adding 30 µL of medium containing 60 nM murine recombinant C5a protein to the left part of the slide.

Techniques: Chemotaxis Assay, Migration