Ohio State University The Effect of BCR and ABL in Hematopoietic Cells Questions Data Analysis Problem – The Effect of BCR/ABL in Hematopoietic Cells Source:

Ohio State University The Effect of BCR and ABL in Hematopoietic Cells Questions

Data Analysis Problem – The Effect of BCR/ABL in Hematopoietic Cells

Source: Tauchi, T., H. S. Boswell, D. Leibowitz, and H. E. Broxmeyer. 1994. Coupling between p210bcr-abl and Shc and Grb2 adaptor proteins in hematopoietic cells permits growth factor receptor-independent link to Ras activation pathway. J. Exp. Med. 179: 167–175. DOI: 10.1084/jem.179.1.167

Experiment

The consequence of the expression of the Bcr/Abl fusion protein in myeloid cell lines was studied in this experiment. The cell lines used were:

• 32D, a non-transformed myeloid cell line

• H7, a mouse myeloid leukemia cell line

• G5, bone marrow cells transformed by a ras oncogene All three cell lines were transfected with an expression vector containing the Bcr/Abl cDNA. Cell extracts were subjected to Western blot analysis using an anti-phosphotyrosine (αP.Tyr) antibody.

Questions

1. Considering the description of the cell lines, explain why there is a difference in band patterns in 32D compared to H7 and G5 cells.

2. What is the consequence of the expression of the Bcr/Abl fusion protein in the three cell lines?

3. What enzyme activity of the Bcr/Abl fusion protein was demonstrated in this experiment?