Extended Data Fig. 8: Miro Reducers Benefit PD Models.
(a) Upper: Typical unfolding curves of human Miro1 protein in the absence (0 μM) and the presence of MR3 are shown. Lower: Quantification of melting temperatures of Miro1 protein in the absence (0 μM) and the presence of MR3 (100 μM). n=4 independent experiments. Two-sided Student T Test. p=0.0000424. (b) α-Syn-A53T-expressing flies (Elav-GAL4>UAS-SNCAA53T) were fed with 10 μM MR5 for 15 days and then their heads were lysed for immunoblotting. The band intensity of DMiro is normalized to that of the mitochondrial loading control VDAC from the same blot. n=4 independent experiments. Two-sided Mann-Whitney Test. p=0.0286. (c) Both Antimycin A and MR5 were dissolved in ethanol. Neurons were pretreated with MR5 24 hrs before the application of Antimycin A for another 6 hrs. The same volume of ethanol was applied at the same time in negative controls. Confocal images overlay triple immunostainings of TH (DA neurons), TUNEL (indicator of cell death), and Dapi (nuclei). The percentage of TUNEL-positive cells out of total cells (Dapi-positive) is calculated in each condition under 20×. n=19 (PD, MR5) or 20 fields (the rest) from 4 independent experiments. p<0.0001. The rest of the precise p values are in Source Data. (d) From images such as in (c), the percentage of TH-positive neurons out of total cells (Dapi-positive) without Antimycin A treatment ranges from 17.08%-19.25%, and is not significantly different among all conditions (p=0.9532), consistent with previous studies from ours and others20,67. n=19 (PD, MR5) or 20 fields (the rest) from 4 independent experiments. (c-d) Comparisons with ‘Wild-type, no treatment’ except otherwise indicated. One-Way Anova Post Hoc Tukey Test. Scale bar: (c) 100 µm. (a) Data are presented as Mean±S.D. with dots. (b-d) Boxes show 25th/75th percentiles, whiskers are the minimum and maximum, and middle line is median.
