Fig. 3

Pathogenic mutations in Parkin RING0 interfere with binding to SLP-2. A Representation of the three-dimensional structure of human Parkin showing the Parkin domains. B Whole cell lysates of SH-SY5Y cells transfected with RING0 Parkin-HA and three mutant RING0 fragments (P153A, K161A and K211A) were subjected to co-IP using an anti-HA antibody, followed by WB analysis of input and IP fractions with the indicated antibodies. Statistical differences were calculated by one-way ANOVA followed by Tukey's post hoc test to correct for multiple comparisons **p ≤ 0.01. C HeLa cells were processed using PLA after transfection with RING0-HA and the mutant RING0 domains using anti-HA and anti-SLP-2 antibodies. For all three mutant RING0 fragments with one of the three missense mutations, a decreased interaction was detected. The PLA signal is visualized in red, DAPI-stained nuclei are shown in blue. Scale bar: 50 µm. Statistical differences were calculated by one-way ANOVA followed by Tukey’s post hoc test to correct for multiple comparisons *p ≤ 0.05, **p ≤ 0.01