Materials and Reagents

1. PC-3 cells (ATCC, CRL-1435, catalog number: 7348669 )
   
2. Three-fold serial dilutions of compounds
   
3. MitoTox^TM MitoBiogenesis^TM In-Cell ELISA Kit (Abcam, catalog number: ab110216 ) including
   
   1. 1,000x IRDye^®-labeled secondary antibodies (species were not disclosed by vendor)
      
   2. 200x primary antibodies (species were not disclosed by vendor)
      
   3. 100x Triton X-100
      
   4. 10x blocking buffer
      
   5. 10x phosphate buffered saline (PBS)
      
   6. 400x Tween-20
      
   7. 1x Janus Green stain (optional)
4. DMSO (cell culture grade) (Sigma-Aldrich, catalog number: D2650 )
   
5. CellTiter Glo (Promega Corporation, catalog number: G7571 )
   
6. Kaighn’s F12 (Life Technologies, Gibco^®, catalog number: 21127 )
   
7. FBS (HyClone, catalog number: SH30071.03 )
   
8. 100 units/ml penicillin and 100 µg/ml streptomycin (P/S) (Life Technologies, Gibco^®, catalog number: 15140-122 )
   
9. Cell fixing reagent: 4% paraformaldehyde (20% paraformaldehyde, VWR International, catalog number: 15713-S )
   
10. Chloramphenicol (10 mM solution in DMSO) (Sigma-Aldrich, catalog number: C1919-5G )
    
11. Kaighn’s F12 medium (see Recipes)
    Note: The cells were maintained in Kaighn’s F12 medium. The cells were passaged twice weekly and maintained at 90% confluence. Cells between passages 4 and 15 were used in the assays.
    

Equipment

1. 96-well plates (collagen I coated 96-well plate) (VWR International, catalog number: 73521-028 )
   
2. Titer plate shaker (Lab-line Instrument, model: 4626 )
   
3. LI-COR^® Odyssey instrument (LI-COR)
   

Software

1. GraphPad Prism 5.0 (GraphPad Software)

Procedure