We analysed the subcellular distribution of p210 BCR-ABL protein using a junction-specific anti-BCR-ABL monoclonal antibody and confocal laser scanning microscopy (CLSM). Our studies have shown that p210 BCR-ABL is arranged in discrete foci in the cytoplasm of cell lines and primary CD34 þ cells but not mononuclear cells suggesting the foci may be a feature of immature chronic myeloid leukaemia cells. We have devised a strategy to score the foci and found the mean number of foci varies between the cell types. The number of foci per cell is directly related to the level of p210 BCR-ABL expression. CLSM was also used to analyse the distribution and colocalization of CT10 regulator-like (CRKL) p210BCR-ABL foci were completely or partially associated, touching or separate in different regions of the same cell. We also analysed the distribution of phosphorylated CRKL (pCRKL) with p210 BCR-ABL and casitas B-cell lymphoma and CRKL but not between p210 BCR-ABL and GRB2. Our observations of the CRKL and p210 BCR-ABL complex may be important for understanding the function of CRKL.
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