Absence of HOXA5 tumor suppressor properties in hematopoietic cells. (A) Typical example of a replating assay with cells transduced with MLLENL and HOXA5 or empty vector as control. (B) Relative levels of total HOXA5 (ie, murine and transduced human HOXA5), human HOXA5, and p53 mRNAs in cells explanted from a replating assay as shown in panel A. (C) Typical example of a replating assay performed with cells cotransduced with HOXA4 and HOXA5 or control vector as indicated. (D) Transcript levels of HOXA4, HOXA5 (total HOXA5), as well as of p53 were determined by qRT-PCR in cells transduced either with HOXA4/HOXA5 or HOXA4/vector as control. To allow a direct comparison of HOXA4 and HOXA5 expression levels, the scale is given in absolute units relative to β-actin (as ΔCt vs β-actin). (E) Homeobox-swap experiments. (Left) HA-specific immunoblot of swap-constructs exchanging the homeobox motif of HOXA5 against the homeobox of HOXA9 (HOXA5homA9) or vice versa (HOXA9homA5). Normal HOXA9 is shown for comparison. (Right) Typical result of a colony-forming assay in which swap-mutants are used.
