Effect of divalent cation on the HK-Mac-1 interaction.

To determine the effect of divalent cations on the HK–Mac-1 interaction, binding assays of FITC–HKa (60 nmol/L) to Mac-1 transfected (open bars) or untransfected (black bars) HEK cells were performed in the presence of different divalent cations as indicated. Cells were preincubated with 10 mmol/L EDTA for 10 minutes at 37°C, then washed with binding buffer without any divalent cation (−mol/L). One of the divalent cations was added to each assay as follows: MgCl₂, MnCl₂, and CaCl₂ at 1 mmol/L, and ZnCl₂ was added at 50 μmol/L.