Rab8a

Protein samples were extracted from cell cultures. Equal amount of protein samples were separated by SDS-PAGE, and transferred to PVDF membranes, which were blocked, and reacted with primary antibodies against Rab8a (1:1000, Abcam, Cambridge, UK), Rab11a (1 : 500, Abcam), or GAPDH (1:1000, Proteintech, Wuhan, China) according to the manufacturer's recommendations. The specific binding of primary antibody was detected by HRP-conjugated species-specific secondary antibody (Beyotime, Shanghai, China) and enhanced chemiluminescence assay.

The following primary antibodies were used: Parkin phospho-Ser65 rabbit monoclonal antibody was raised by Epitomics/Abcam in collaboration with the Michael J. Fox Foundation for Research (please contact tools@michaeljfox.org for questions). Ubiquitin phospho-Ser65 (Cell Signalling Technology (CST)), OPA1 (CST), PINK1 (Novus), PINK1 (in-house generated by Dundee Cell Products (DCP)), Parkin (Santa Cruz), Rab8A (Abcam), Rab8A phospho-Ser111 (Abcam), GAPDH (Santa Cruz) and Vinculin (CST). The polyclonal phospho-Ser228 PINK1 was generated by the Michael J. Fox Foundation's research tools program in partnership with Abcam (development of a monoclonal antibody is under way; please contact tools@michaeljfox.org for questions). The following antibody was produced by the MRCPPU Reagents and Services at the University of Dundee in sheep: anti-PINK1 (S085D).