BCR-ABL induces gene expression levels of SOCS proteins.

<p><b>A.</b> Ba/F3 cells were transduced with BCR-ABL or an empty vector control. Prior to RNA extraction, cells were starved over night from IL-3. RT-PCR was performed for SOCS family members. Relative gene expression normalized to <i>RNA pol II</i> is depicted. The experiment was performed three times, the error bars are indicating the standard deviation. <b>B.</b> Sca-1+ cells were enriched from murine BM and transduced with BCR-ABL or an empty vector control. GFP-sorted cells were starved over night from cytokines prior to RNA extraction. Relative gene expression of SOCS family members is shown. Relative gene expression normalized to <i>RNA pol II</i> is depicted. The experiment was performed three times, the error bars are indicating the standard deviation. *P ≤ 0.05, **P ≤ 0.009, ***P ≤ 0.001.</p

Mouse transplantation model.

<p>Transduction efficiency of Sca-1⁺ cells was normalized according to GFP expression and 2.5 x 10⁴ GFP-positive cells were transplanted into the tail vein of sub-lethally irradiated recipient mice. Moribund mice were sacrificed and analyzed. All data shown are from time of death. <b>A.</b> Disease free survival of transplanted animals is plotted in Kaplan-Meier curves. <b>B.</b> White blood counts and hemoglobin content in the peripheral blood. <b>C.</b> Spleen and liver weights of analyzed mice. <b>D.</b> Spleen samples were stained with H&E. Representative images are shown.</p

Gene expression levels of SOCS proteins after BCR-ABL kinase inhibition.

<p><b>A-B.</b> Philadelphia chromosome positive (Ph+) or negative (Ph-) long term cultures of primary human acute lymphoblastic leukemia cells were treated with 1 μM imatinib for 16h. RNA was extracted and RT-PCR was performed for SOCS family members. Relative gene expression normalized to B2M is shown. The experiment was performed three times, the error bars are indicating the standard deviation. **P ≤ 0.009, ***P ≤ 0.001 <b>C.</b> K562 cells were treated with 2 μM imatinib for 16h. RNA was extracted and RT-PCR performed for SOCS family members. Gene expression relative to B2M is depicted. The experiment was performed three times, the error bars are indicating the standard deviation. *P ≤ 0.02, **P ≤ 0.009, ***P ≤ 0.001 <b>D.</b> K562 cells were washed with PBS and resuspended in RPMI medium containing 1% FCS. BCR-ABL was inhibited for 16h with 2 μM imatinib or 20 nM dasatinib. Total cell lysates were immunoblotted and probed with indicated antibodies. Tubulin is shown as loading control. Inhibition of BCR-ABL was assessed by its phosphorylation status. The experiment was performed three times.</p