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. 2000 Nov 14;97(24):13354–13359. doi: 10.1073/pnas.240347797

Figure 2.

Figure 2

UbcH8 interacts preferentially with the C-terminal R2 ring-finger domain of Parkin. (A) Schematic representation of putative functional domains of Parkin used in the mapping experiments. (B) Lysates prepared from HEK 293 cells transfected with pRK5-HA-UbcH8 and various Myc-tagged Parkin domain constructs were subjected to immunoprecipitation (IP) with anti-myc antibody followed by anti-HA immunoblotting (Upper). The blot was stripped and reprobed with the anti-myc antibody (Lower) to illustrate that equivalent levels of the Parkin constructs were present in the extracts. This experiment was replicated three times with similar results. (C) Lysates prepared from HEK 293 cells transfected with pRK5-HA-UbcH8 and various Myc-tagged Parkin mutant constructs (see text for details) were subjected to immunoprecipitation (IP) with anti-myc antibody followed by anti-HA immunoblotting (Upper). The blot was stripped and reprobed with the anti-Myc antibody (Lower) to illustrate that equivalent levels of the Parkin constructs were present in the extracts. This experiment was replicated three times with similar results.