FPR2/ALX-Expressing HL60 Cell Calcium Flux

HL60 cells stably transfected with human FPR2/ALX have been recently described (5 (link)). These cells were grown in RPMI medium (Biochrom) supplemented with 10% FCS (Sigma-Aldrich), 20 mM HEPES (Biochrom), penicillin (100 units/ml), streptomycin (100 µg/ml) (Gibco), 1× GlutaMAX (Gibco), and G418 (Biochrom) at a final concentration of 1 mg/ml. Calcium fluxes were analyzed by stimulating cells loaded with Fluo-3-AM (Molecular Probes), and the fluorescence was monitored with a FACSCalibur flow cytometer (Becton, Dickinson), as recently described (39 (link)).

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Schlatterer K., Beck C., Hanzelmann D., Lebtig M., Fehrenbacher B., Schaller M., Ebner P., Nega M., Otto M., Kretschmer D, & Peschel A. (2018). The Mechanism behind Bacterial Lipoprotein Release: Phenol-Soluble Modulins Mediate Toll-Like Receptor 2 Activation via Extracellular Vesicle Release from Staphylococcus aureus. mBio, 9(6), e01851-18.

Publication 2018

RPMI medium FCS HEPES penicillin streptomycin GlutaMAX Fluo-3-AM FACSCalibur flow cytometer

Calcium Cells Fluo 3 Fluorescence G418 Hepes Hl60 cells Human Molecular probes Penicillin Streptomycin

Corresponding Organization :

Other organizations : University of Tübingen, University Children's Hospital Tübingen, National Institute of Allergy and Infectious Diseases, National Institutes of Health

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Variable analysis

independent variables

Stimulating cells loaded with Fluo-3-AM

dependent variables

Calcium fluxes

control variables

RPMI medium (Biochrom) supplemented with 10% FCS (Sigma-Aldrich), 20 mM HEPES (Biochrom), penicillin (100 units/ml), streptomycin (100 µg/ml) (Gibco), 1× GlutaMAX (Gibco), and G418 (Biochrom) at a final concentration of 1 mg/ml

controls

No positive or negative controls were explicitly mentioned.

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