Fig 1: ML335 attenuates mitochondrial dysfunction and suppresses mitochondrial fission via down-regulating March5. (A) Western blotting was used to detect the reverse effect of March5 overexpression on Ac-SOD2 protein expression inhibited by ML335. (B) MitoSOX probe staining and flow cytometry assay were used to detect mitochondrial ROS. The relative level of MitoSOX was displayed on the right. (C) Mitochondrial membrane potential was measured by flow cytometry after staining with the JC-1 probe. Statistical analyses of the proportion of cells with low mitochondrial membrane potential are shown on the right. (D) Western blotting was used to detect the effect of ML335 on p-LKB1 and LKB1 protein levels. The relative level of p-LKB1/LKB1 (Vinculin as loading control) was displayed on the right. (E) March5 overexpression prevented the up-regulation of p-LKB1 by ML335 as detected by Western blotting. The relative level of p-LKB1/LKB1 (Vinculin as loading control) was displayed on the right. (F) The effect of ML335 on p-AMPK detected by Western blotting. The relative level of p-AMPK/AMPK (Vinculin as loading control) was displayed on the right. (G) LKB1 knockdown prevented the upregulation of p-AMPK protein by ML336 as detected by Western blotting. The relative level of p-AMPK/AMPK (Vinculin as loading control) was displayed on the right. (H, I) Compound C reversed the down-regulation of MFF and p-DRP1 by ML335 as detected by Western blotting. The relative level of p-AMPK/AMPK (Vinculin as loading control) was displayed on the right