2.11. Measurement of the labile iron pool (LIP)

LIP was measured as described previously. In brief, primary hepatocytes were incubated with 0.05 μM calcein acetoxymethyl ester (Sigma-Aldrich, 17783) for 30 min at 37 °C. The cells were then washed with PBS and incubated with deferiprone (DFO, 100 μM) for 1 h at 37 °C or left untreated. The cells were analysed using a flow cytometer. The difference in the mean cellular fluorescence with and without DFO incubation reflected the level of LIP.