Subcellular distribution of p210BCR-ABL in CML cell lines and primary CD34+ CML cells

Abstract

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.

Non-receptor Tyrosine Kinases c-Abl and Arg Regulate the Activity of C/EBPβ

“…The phosphorylation may be mediated by the P145 isoform of Abl, similar to c-Abl, which was found distributed in the nucleus and in the cytoplasm. 38 Our findings that the nuclear P145 isoform of Abl in K562 cells formed a complex with C/EBPβ in nuclear extracts also support the hypothesis.…”

PYK2 sustains endosomal-derived receptor signalling and enhances epithelial-to-mesenchymal transition

“…Co-localization was measured using the Zeiss LSM 510 meta software, essentially as previously described 64 . In brief, the overlapping regions between the green and red channels, which appeared in yellow in the merged confocal images, were marked as regions of interest in randomly selected cells (n420).…”

Antibody arrays identify protein‐protein interactions in chronic myeloid leukaemia

“…This antibody binds specifically to p210 BCR-ABL1 with the b2a2 (e13a2) breakpoint. We have previously used this antibody to analyse the subcellular distribution of p210 BCR-ABL1 by confocal microscopy (Patel et al, 2008). The antibody also immunoprecipitates p210 BCR-ABL1 from cell lines with the b2a2 breakpoint but not from K562 cells (b3a2 breakpoint) (data not shown).…”