Primary nasal epithelial cells were obtained by digesting nasal turbinates or polyps with 0.5 mg/mL collagenase IV (Worthington Biochemical Corp., USA) for 1 hour in Hanks’ balanced salt solution (HBSS; Sigma-Aldrich, NL) followed by an incubation with anti-EpCAM MicroBeads (Miltenyi Biotec, DE) and a positive selection on a magnetic column. Epithelial cells were grown in 75 mL flasks in BEGM growth medium (Lonza Clonetics, NL). Culture medium was replaced every other day. Cells were grown in fully humidified air containing 5% CO2 at 37°C.
The human airway epithelial cell line NCI-H292 (ATCC, USA) was cultured in RPMI 1640 medium supplemented with 10% (v/v) fetal calf serum (HyClone, USA), 1.25 mM of glutamine, 100 U/mL of penicillin, and 100 μg/mL of streptomycin. Creation of the EGR-1 and DUSP-1 knock-down mutants is described elsewhere [18 (link)].
The human airway epithelial cell line NCI-H292 (ATCC, USA) was cultured in RPMI 1640 medium supplemented with 10% (v/v) fetal calf serum (HyClone, USA), 1.25 mM of glutamine, 100 U/mL of penicillin, and 100 μg/mL of streptomycin. Creation of the EGR-1 and DUSP-1 knock-down mutants is described elsewhere [18 (link)].