Figure 5. Functional interaction of Fbxo7 with PINK1.

a, PINK1 localisation at the mitochondria was assessed by immunocytochemistry in SH-SY5Y cells transfected with PINK1-HA plus scrambled (scr) or Fbxo7 siRNA following 1 or 3 h treatment with CCCP (10 μM). Cells were scored visually for the co-localisation of PINK1-HA with complex V β subunit (CxVβ), a mitochondrial marker. Histograms indicate the percentage of cells in which PINK1-HA accumulated at the mitochondria. Data are presented as mean ± S.E.M., * p < 0.05. Representative images are displayed for cells transfected as indicated, following 0 or 3 h CCCP treatment. For corresponding images at 0 h and 1 h treatment, see Supplementary Figure 5a. Scale bar, 10 μm. b, Fbxo7 accumulation in the mitochondrial fraction following treatment with CCCP (10 μM) is impaired in SH-SY5Y cells transfected with PINK1 siRNA compared to scrambled siRNA (scr). Full-length blots are presented in Supplementary Figure S9. c-f, Overexpression of Fbxo7 does not rescue (c, e) climbing or (d, f) flight defects in PINK1 male mutants (PINK1^B9) or PINK1:parkin double mutants (PINK1^B9;park²⁵, daG4). Control genotype is (c, d) PINK1^B9/+;da-GAL4/+ and (e, f) da-GAL4/+. Histograms indicate mean ± S.E.M. Significance was determined by one-way ANOVA with Bonferroni correction (*** p < 0.001). For climbing and flight assays at least 50 flies were assessed.