American Society of Hematology

Figure 5

$Figure 5. E2F3 is a bona fide target of BCR/ABL tyrosine kinase activity. (A) Western blot shows E2F3a (top arrow) and E2F3b (bottom arrow) protein expression in CD34+ BMCs from a healthy donor (NBM), patients with CML-CP, and patients with CML-BC (n = 3 per group). (B) E2F3 protein levels in the CML-BC (n = 2) CD34− and in untreated or imatinib-treated (for the indicated times) CD34+ BMC fractions. GRB2 was used as a control for equal loading. *Nonspecific band detected by a new lot of the E2F3 antibody. (C) Growth factor–independent methylcellulose colony formation of 32D-BCR/ABL cells transduced with the empty pSUPER.retro.neo + GFP retrovirus (), or with retroviruses carrying shRNA for E2F3a () or E2F3a/b (). Inset: Western blot shows shRNA-mediated E2F3a/b down-regulation. (D) Analysis of tumors from SCID mice subcutaneously injected with vector-transduced () or E2F3a/b shRNA–transduced () 3 2D-BCR/ABL cells.$

E2F3 is a bona fide target of BCR/ABL tyrosine kinase activity. (A) Western blot shows E2F3a (top arrow) and E2F3b (bottom arrow) protein expression in CD34⁺ BMCs from a healthy donor (NBM), patients with CML-CP, and patients with CML-BC (n = 3 per group). (B) E2F3 protein levels in the CML-BC (n = 2) CD34⁻ and in untreated or imatinib-treated (for the indicated times) CD34⁺ BMC fractions. GRB2 was used as a control for equal loading. *Nonspecific band detected by a new lot of the E2F3 antibody. (C) Growth factor–independent methylcellulose colony formation of 32D-BCR/ABL cells transduced with the empty pSUPER.retro.neo + GFP retrovirus (), or with retroviruses carrying shRNA for E2F3a () or E2F3a/b (). Inset: Western blot shows shRNA-mediated E2F3a/b down-regulation. (D) Analysis of tumors from SCID mice subcutaneously injected with vector-transduced () or E2F3a/b shRNA–transduced () 3 2D-BCR/ABL cells.

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