FIG. 8.
Pak5 is present in the mitochondrial fraction. A cellular fractionation was performed on CHO stable cell lines as described in Materials and Methods to assess Pak5 presence in the cellular compartments. (A) Equal amount of proteins from cytosolic (Cy) S100, light microsomes (LM) and mitochondrial (M) fractions were loaded on a gel, and Western blotting was performed using anti-Myc antibody. The mitochondrial fraction purity was assessed using an anti-COX IV and anticalnexin antibodies. The fractions were also subjected to immunoblotting with anti-BAD, anti-BCL-2 and anti-BCL-xL antibodies. (B) CHO cells expressing vector alone, WT Pak5, or KD Pak5 were treated with CA (10 μM) or C2C (50 μM) for 16 h. A cellular fractionation was then performed. Equal amount of proteins from cytosolic (Cy) S100, light microsomes (LM) and mitochondrial (M) fractions were loaded on a gel, and Western blotting was performed using anti-Myc antibody. The mitochondrial fraction purity was assessed using an anti-COX IV and anticalnexin antibodies. (C) CHO cells were transfected with pCMV6, HA-Pak4 or HA-Pak6, and a subcellular fractionation was performed 48 h after transfection as described above.