Figure 2. Miro1 Reducer Eliminates the Miro1 Defect in PD Fibroblasts.

(A) Chemical properties of Miro1 Reducer. (B) Wild-type flies (w¹¹¹⁸) were fed with DMSO alone (0 μM Miro1 Reducer) or Miro1 Reducer in DMSO at 250 μM for 7 days, and lysed and blotted as indicated. The band intensities are normalized to those of Tubulin from the same blots. n=4. (C-E) Fibroblasts were treated, lysed, and blotted as indicated. (C) Band intensities are normalized to those of GAPDH from the same blots and compared to “Healthy-1, no treatment” except otherwise indicated. n=4. (D) Band intensities are normalized to those of ATP5β from the same blots and expressed as a percentage of the mean of “0 μM Miro1 Reducer with CCCP”. Mean±S.E.M is shown. n=3. (E) Band intensities are normalized to those of GAPDH from the same blots and compared to “Healthy-1, no CCCP, no MG132” except otherwise indicated. n=4. (F) Healthy-1 and PD-2 fibroblasts were transfected as indicated, and immunostained with anti-Myc (Miro1, green) and stained with Dapi (blue). The Miro1 intensity is normalized to that of mito-dsRed from the same cell and quantified across 37-62 cells from 5 fields each transfection, 3 independent transfections. Scale bars: 50 μm. For (C-F), both CCCP and Miro1 Reducer were dissolved in DMSO. Cells were pretreated with Miro1 Reducer 24 hrs before the application of CCCP for another 6 hrs. The same volume of DMSO was applied at the same time in negative controls. Cell line information is in Table S1. See also Figure S3–S4.