Quantification of lipids
phospholipidosis, Steatosis
PHOSPHOLIPIDOSIS, STEATOSIS
Lipid accumulation was measured using HCS LipidTOX Phospholipidosis and Steatosis Detection Kit according to the manufacturer’s instructions (ThermoFisher, USA; H34158). Briefly, cells were incubated in complete bronchial epithelial growth medium supplemented with 1 x phospholipidosis detection reagent for 48 hr. Cells were subsequently fixed in 4% PFA and stained with 1 X neutral lipid detected reagent for 30 min and counterstained with 1 μg mL-1 Hoechst 33258. Staining intensity was normalized to the amount of Hoechst 33258 positive nuclei across multiple fields of view.