Figure S2.
Parkin-mediated Miro degradation correlates with the dissociation of VPS13D from mitochondria. COS7 or HeLa cells were cotransfected with myc-Miro1, VPS13D, and EGFP-Parkin. (A and D) Western blots showing the decrease of myc-Miro1 level upon 10 µM valinomycin treatment of COS7 (A) or HeLa cells (D). Scarlet, a modified RFP, was visualized by anti-RFP antibodies. Sizes in kD are indicated next to each blot. (B, C, and E) Confocal time lapses of COS7 (B and C) or HeLa cells (E) coexpressing EGFP-Parkin, VPS13D^Halo, and myc-Miro1. VPS13D, initially recruited to mitochondria by Miro, is shed from mitochondria and relocated to the cytosol upon treatment with valinomycin, which induces the recruitment of EGFP-Parkin. The boxed regions in B are shown in higher magnification in C. Scale bars, 30 µm in the main panels and 3 µm in insets.

Parkin-mediated Miro degradation correlates with the dissociation of VPS13D from mitochondria. COS7 or HeLa cells were cotransfected with myc-Miro1, VPS13D, and EGFP-Parkin. (A and D) Western blots showing the decrease of myc-Miro1 level upon 10 µM valinomycin treatment of COS7 (A) or HeLa cells (D). Scarlet, a modified RFP, was visualized by anti-RFP antibodies. Sizes in kD are indicated next to each blot. (B, C, and E) Confocal time lapses of COS7 (B and C) or HeLa cells (E) coexpressing EGFP-Parkin, VPS13D^Halo, and myc-Miro1. VPS13D, initially recruited to mitochondria by Miro, is shed from mitochondria and relocated to the cytosol upon treatment with valinomycin, which induces the recruitment of EGFP-Parkin. The boxed regions in B are shown in higher magnification in C. Scale bars, 30 µm in the main panels and 3 µm in insets.

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