Expression of TEL-AML1 in murine hematopoietic progenitor cells. (A) Schematic diagram of the MSCV constructs used to express TEL-AML1(HA) and EGFP in hematopoietic progenitor cells. E indicates EcoRI; N, NotI. (B) Southern blot analysis of genomic DNA from retrovirally transduced 3T3 cells demonstrating integration of the MSCV-T/A provirus. The indicated cDNA fragment (A) was used as a probe to detect the 2.4-kilobase proviral band (indicated by an arrow) following EcoRI/Not I digestion of genomic DNA. (C) Western blot analysis of protein extracts from 3T3 cells transduced with MSCV-T/A or MSCV-EGFP. Anti-TEL, anti-AML1, or anti-HA antibodies detect TEL-AML1(HA) as a doublet of 95 kDa, due to alternative TEL translational start sites.33 (D) Flow cytometric analysis of HPCs 48 hours following infection with concentrated viral supernatants. Gray lines represent uninfected cells, black lines depict transduced cells, and numbers refer to the percentage of cells transduced. (E) Detection of TEL-AML1(HA) protein in HPCs 48 hours after infection, with anti-HA antibody by Western blot analysis.
