FIG. 2.

Subcellular distribution of c-Abl. (A) ER, cytoplasmic (Cyto), and mitochondrial (Mito) fractions were isolated from Rat1 cells. Equal amounts of protein (5 μg) from each fraction were subjected to immunoblotting (IB) with anti-c-Abl, anticalreticulin, anti-β-actin, or anti-HSP60. (B) Rat1 cells (2 × 10⁷) were divided into five aliquots for preparation of total cell, nuclear, cytoplasmic, ER, and mitochondrial lysates. The lysates were adjusted to 500 μl with PBS, and aliquots (20 μl) were subjected to immunoblotting with anti-c-Abl. Signal intensities were analyzed by densitometric scanning. The results are presented as the percentage of c-Abl in each subcellular fraction compared to that in the total cell lysate. (C) ER and plasma membrane preparations were isolated from Rat1 cells. Equal amounts of protein (5 μg) were subjected to immunoblot analysis with anti-c-Abl, anticalreticulin, and anti-PDGF-R.