Validation and expression analysis of the hnRNP-A1–associated mRNAs in BCR/ABL cell lines and patient-derived CML BMCs. (A) RT-PCR for cyclin D2, HOXB7, SET, SNF2β, ILF3, Tra2β, E2F3a, and E2F3b on hnRNP-A1–immunoprecipitated mRNA from K562-A1-HA cytoplasmic lysates. hnRNP-A1 RNP complexes isolated from K562 cytoplasmic fractions immunoprecipitated with an anti–hnRNP-A1 antibody (9H10; lane 3) or an unrelated isotype-matched anti-FLAG antibody (clone M2; lane 2). mRNA extracted from mRNP-enriched lysates was used as a positive control (lane 1). (B) Western blots show E2F3, SNF2β, and Tra2β expression in untreated and imatinib-treated K562 cells, doxycycline-induced and uninduced TonB2.10 cells, and in parental and untreated or imatinib-treated BCR/ABL-expressing 32Dcl3 cells. GRB2 levels were detected as a control for equal loading. Right panels: Real-time PCR shows expression of ILF3 and HOXB7 mRNA in untreated and imatinib-treated BCR/ABL-expressing myeloid cells and in the BaF3-derived BCR/ABL-inducible TonB2.10 lymphoid cell line (expressed as mean ± standard error). GAPDH was used for normalization. (C) SNF2β, E2F3, Tra2β, and GRB2 protein levels and BCR/ABL phosphorylation in mononuclear BMCs from 2 patients with CML-CP and CML-BC. (D) Effect of shRNA-mediated hnRNP-A1 downmodulation on hnRNP-A1, SET, SNF2β, and E2F3 protein levels.