Fig. 4

Arnidiol induces mitochondrial translocation of Drp1 and Cofilin. a and d MDA-MB-231 cells were treated with various concentrations of Arn for 48 h or with Arn (60 μM) for different time intervals as indicated, cytosol and mitochondrial fractions were prepared and subjected to western blot using antibodies against Drp1 and Cofilin. GAPDH and COX IV were used as loading controls. b and e MDA-MB-231 cells were treated with Arn (60 μM) for 48 h, the colocalization of MitoTracker (red) and Drp1 (green) or Cofilin (green) was examined using confocal microscopy. Scale bars: 10 μm. c and f MCF-7, Eca109, SMMC-7721 and A549 cells were treated with Arn (60 μM) for 48 h, cytosol and mitochondrial fractions were prepared and subjected to western blot using antibodies against Drp1 and Cofilin. GAPDH and COX IV were used as loading controls. g MDA-MB-231 cells were treated with Arn (60 μM) for 48 h, WCL, mitochondrial and cytosol fractions were prepared and subjected to immunoprecipitation using anti-Cofilin, the associated Cofilin and Drp1 were determined using immunoblotting. h MDA-MB-231 cells were treated with Arn (60 μM) for 48 h, the colocalization of Cofilin (red), Drp1 (green), and MitoTracker (blue) was examined using confocal microscopy. Scale bars: 10 μm