flow cytometry tubes with cell strainer snap cap  (Corning Life Sciences)

Journal: Cell Communication and Signaling : CCS

Article Title: F0F1 ATP synthase regulates extracellular calcium influx in human neutrophils by interacting with Ca v 2.3 and modulates neutrophil accumulation in the lipopolysaccharide-challenged lung

doi: 10.1186/s12964-020-0515-3

Figure Lengend Snippet: F-ATPase/Ca v 2.3 functional complexes regulate ERK 1/2 phosphorylation and ROS production after fMLP activation of neutrophils. a. A representative western blot is presented to show the effect of F-ATPase/Ca v 2.3 functional complexes on ERK 1/2 phosphorylation. Neutrophils were pretreated with 400 nM SNX482 and 50 μg/ml oligomycin A in Ca 2+ -containing HBSS for 30 min, followed by 1 min of 100 nM fMLP activation. Cells in Ca 2+ -containing HBSS or Ca 2+ -free HBSS incubated with or without fMLP were used as positive or negative controls, respectively. b. The effect of F-ATPase/Ca v 2.3 functional complexes on ROS production was assessed by flow cytometry. DHR123-loaded neutrophils were pretreated with 400 nM SNX482 and 50 μg/ml oligomycin A in Ca 2+ -containing HBSS for 30 min, followed by 30 min of 100 nM fMLP incubation. Cells in Ca 2+ -containing HBSS or Ca 2+ -free HBSS incubated with or without fMLP were used as positive or negative controls, respectively

Article Snippet: The aggregated cells were removed by filtration through a 35-μm cell strainer snap cap on a flow cytometry tube (Falcon, Corning, Cambridge, MA).

Techniques: Functional Assay, Activation Assay, Western Blot, Incubation, Flow Cytometry