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. 2016 Jun 22;35(15):1656–1676. doi: 10.15252/embj.201694401

Figure EV3. C9orf72 regulates translocation of the ULK1 complex to the phagophore via Rab1a.

Figure EV3

  1. HeLa cells were co‐transfected with empty vector (EV), FLAG‐C9orf72S or FLAG‐C9orf72S (green), Myc‐Rab1aWT or dominant negative Myc‐Rab1aS25N (DN), and mCherry‐FIP200 (red). 24 h post‐transfection, cells were treated with Torin1 (250 nM; 3 h). Translocation of the ULK1 complex was quantified as the number of mCherry‐FIP200‐positive puncta per cell from 7 independent experiments (mean ± SEM; one‐way ANOVA with Fisher's LSD test: ns, not significant, **P ≤ 0.01, ****P ≤ 0.0001; N (cells) = WT/EV: 142; WT/EV/Torin1: 73; WT/C9orf72L: 108; WT/C9orf72S: 123; DN/EV: 159; DN/EV/Torin1: 150; DN/C9orf72L: 159; DN/C9orf72S: 123). Scale bar = 10 μm.
  2. HeLa cells were co‐transfected with empty vector (EV), FLAG‐C9orf72S or FLAG‐C9orf72S (red), Myc‐Rab1aWT or dominant negative Myc‐Rab1aS25N (DN), and EGFP‐LC3 (green). 24 h post‐transfection, cells were treated with Torin1 (250 nM; 3 h). Autophagosomes were quantified as the number of EGFP‐LC3‐positive puncta per cell from 2 independent experiments (mean ± SEM; one‐way ANOVA with Fisher's LSD test: ns, not significant, **P ≤ 0.01, ****P ≤ 0.0001; N (cells) = WT/EV: 51; WT/EV/Torin1: 53; WT/C9orf72L: 56; WT/C9orf72S: 41; DN/EV: 53; DN/EV/Torin1: 53; DN/C9orf72L: 55; DN/C9orf72S: 42). Scale bar = 10 μm.