Fig. 6.

NPC cells show mitochondrial morphological alterations and MLN64 down-regulation improves mitochondrial function in NPC cells. CHO Wild-type (WT) and Npc1^-/- (NPC) cells were transfected with nontargeting siRNA (siNT) and a siRNA directed against MLN64 (siMLN64) for 96 h. (A) 30 μg of proteins from cell homogenates were subjected to SDS-PAGE and analyzed by western blot using antibodies against the fission (FIS1) and fusion (MFN-2, OPA-1) related proteins . (B) The FIS1 bands intensities were measured using ImageJ and normalized with the ε-Cop protein. *p<0.05 (n=3) (C) Cells were immunostained with anti-MLN64 antibody (red), mitotracker green and nuclei were stained with Hoechst. (D) Cells were stained using Mitotracker red to measure the MMP or Mitosox (E) to quantify mitochondrial superoxide production 96 h post transfection. ^aStatistically different from WT. ^bStatistically different from NPC. ^cStatistically different from NPC+siNT. (n=3). Scale bar: 10 µm, zoom 5 µm.