Figure 2. WT Parkin complements the mislocalization of a catalytically inactive Parkin mutant in trans. (A) HeLa cells expressing GFP-Parkin(C431S) (catalytically inactive mutant) with or without WT HA-Parkin were treated with CCCP (10 µM, 1.5 h), and immunostained with anti-GFP and anti-Tom20 antibodies. (B) Coexpression of HA-Parkin(C431S) did not complement the localization defect of GFP-Parkin(C431S). (C) The number of cells with GFP-Parkin(C431S) localized to the mitochondria was counted in 100 cells. Numbers on the horizontal axis correspond to those of A. Bars represent the mean ± SD values of three experiments (error bars). (D) Mitochondrial localization of Mt-Parkin (Parkin fused with a MitoNEET mitochondrial outer membrane–targeting signal). (E) Cells expressing GFP-Parkin(C431S) (catalytically inactive mutant) with or without Mt-Parkin were treated with CCCP (10 µM, 1 h), and immunostained with anti-GFP and anti-Tom20 antibodies. (F) The number of cells with GFP-Parkin(C431S) localized to the mitochondria was counted as in C. Numbers on the horizontal axis correspond to those of E. Bars, 10 µm.
Figure 2.

WT Parkin complements the mislocalization of a catalytically inactive Parkin mutant in trans. (A) HeLa cells expressing GFP-Parkin(C431S) (catalytically inactive mutant) with or without WT HA-Parkin were treated with CCCP (10 µM, 1.5 h), and immunostained with anti-GFP and anti-Tom20 antibodies. (B) Coexpression of HA-Parkin(C431S) did not complement the localization defect of GFP-Parkin(C431S). (C) The number of cells with GFP-Parkin(C431S) localized to the mitochondria was counted in 100 cells. Numbers on the horizontal axis correspond to those of A. Bars represent the mean ± SD values of three experiments (error bars). (D) Mitochondrial localization of Mt-Parkin (Parkin fused with a MitoNEET mitochondrial outer membrane–targeting signal). (E) Cells expressing GFP-Parkin(C431S) (catalytically inactive mutant) with or without Mt-Parkin were treated with CCCP (10 µM, 1 h), and immunostained with anti-GFP and anti-Tom20 antibodies. (F) The number of cells with GFP-Parkin(C431S) localized to the mitochondria was counted as in C. Numbers on the horizontal axis correspond to those of E. Bars, 10 µm.

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