Figure 5
From: GBF1 and Arf1 interact with Miro and regulate mitochondrial positioning within cells
Miro GTPases interact physically with GBF1. (A,B) RPE1 cells were cotransfected with Venus or Venus-tagged GBF1 and either Myc or Myc-Miro1 (A) or Myc-Miro2 (B). Immunoprecipitations were carried out with GBP-beads. Cells lysates (Input) and immunoprecipitated proteins were analyzed by Western blotting using anti-GFP and anti-Myc antibodies. The input lanes represent 15% of lysate used in immunoprecipitation reactions. (C) RPE1 cells were cotransfected with Arf1-T31N-GFP, Arf1-Q71L-GFP or GFP and Myc-Miro2. GFP-tagged proteins were immunoprecipitated with magnetic GFP-Trap®_A. Cells lysates (Input) and immunoprecipitated proteins were analyzed by Western blotting using GFP- and Myc-antibodies. The input lanes represent 10% of lysate used in immunoprecipitation reactions. One representative experiment of three independent experiments is shown. Bands were quantified by a densitometric analysis performed using Image J software. Results are the mean of the ratio between the Myc and the GFP band values ± S.D. of the three independent experiments. *P ≤ 0.05 versus control cells.
