Fig. 4

Mitochondrial fraction of LRRK2 and GTP binding activity of MFN2 were increased upon fMLP stimulation. A-C The mitochondrial fraction of LRRK2 was increased upon fMLP stimulation. dHL-60 cells expressing tetracycline-inducible Lrrk2 shRNA or irrelevant (Irr) shRNA were cultured in 1.25% DMSO with 0.5 µg/ml doxycycline (Dox) for 3 days. The mitochondrial fraction of LRRK2 and MFN2 were measured with or without 1 µM fMLP for 5 min. D, E Lrrk2 knockdown results in a decrease in MFN2 GTP-binding activity of dHL-60 cells. dHL-60 cells expressing tetracycline-inducible Lrrk2 shRNA or irrelevant (Irr) shRNA were cultured in 1.25% DMSO with 0.5 µg/ml  Dox for 3 days. The GTP-binding activity of MFN2 was measured with or without 1 µM fMLP for 5 min. A, D Representative western blotting images. B The rate of LRRK2/TOMM20 in the mitochondrial fraction of Irr shRNA expressing dHL-60 cells without fMLP stimulation was taken as 1.0 (n = 3). ** p < 0.01 compared with Irr shRNA, Lrrk2 sh1, or sh2 expressing dHL-60 cells without fMLP. C The rate of MFN2/TOMM20 in the mitochondrial fraction of Irr shRNA expressing dHL-60 cells without fMLP stimulation was taken as 1.0 (n = 3). NS, not significant. E GTP-binding activity of Irr shRNA expressing dHL-60 cells without fMLP was taken as 1.0 (n = 4). Error bars, SEM. ** p < 0.01 and * p < 0.05 compared with Irr shRNA expressing dHL-60 cells with or without 1 µM fMLP. NS, not significant