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Mitochondrial expression of phospho-mimetic polyUb chain improves mitochondrial degeneration by PINK1 inactivation in Drosophila.

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posted on 04.12.2014, 15:10 by Kahori Shiba-Fukushima, Taku Arano, Gen Matsumoto, Tsuyoshi Inoshita, Shigeharu Yoshida, Yasushi Ishihama, Kwon-Yul Ryu, Nobuyuki Nukina, Nobutaka Hattori, Yuzuru Imai

(A) Tom70MTS-2FLAG-4xUb (WT, SA or SE) expression alone does not alter the mitochondrial morphology of thorax IFMs in Drosophila. To visualise the mitochondria, the mitoGFP (green) transgene was co-expressed, and the muscle tissue was counterstained with phalloidin (magenta). (B) TEM images of the IFMs in the indicated genotypes of 7-day-old adult flies are shown. (C) Tom70MTS-2FLAG-4xUb (WT, SA or SE) or LacZ was expressed in the thorax muscle of PINK1-deficient flies using the MHC driver. dMfn levels from the thoraxes of 3-day-old adult flies were analyzed by western blot. Actin served as a loading control. (D) The band intensities of dMfn were normalized to each actin signal shown in (C). The values (arbitrary units; A.U.) represent the mean ±SEM of five independent samples as in (C). ** p<0.01 vs. PINK1−/−; LacZ, § p<0.05. (E) TEM images of the IFMs in 3-day-old Parkin-deficient flies expressing LacZ or Tom70MTS-2FLAG-4xUb SE. Scale bars  = 5 µm in (A), 2 µm in (B, upper), 1 µm in (B, lower and E).