Western blot analysis

Cells were plated in T25 flasks and treated when 80% confluent for 24 hours with 20 μM of each compound (honokiol, honokiol DCA, and hexafluoro. Blots were probed with antibodies for p-akt, p-mapk 42/44, p-p38, p-DRP1, total akt, total ERK, FOXD3, MDMX, p-Rac1b, MnSOD or beta actin were added at a concentration of 1:1000 in 5% non-fat dry milk in TBS and allowed to shake overnight in the coldroom (4°C) [48]. The next day, the blots were probed with anti-rabbit or anti-mouse HRP linked antibodies 1:10,000 (Cell Signaling) 5% non-fat dry milk in TBS with anti-rabbit or anti-mouse HRP linked antibodies (Cell Signaling). Super-Substrate from Thermofisher was used to activate the HRP linked secondary antibodies for development. The membranes were then developed using a Bio-rad docking station and camera. The software used was Bio-Rad Image Lab version 4.0.