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Fig. 6 | Journal of Hematology & Oncology

Fig. 6

From: CRISPR/CAS9-mediated knockout of Abi1 inhibits p185Bcr-Abl-induced leukemogenesis and signal transduction to ERK and PI3K/Akt pathways

Fig. 6

Effect of imatinib treatment on IL3-independent growth and leukemogenesis of Abi1-deficient p185Bcr-Abl cells. a. Imatinib induces dose-dependent cell death in control p185Bcr-Abl cells (p185 Cas9) and Abi1-deficient p185Bcr-Abl cells (p185 KO2.3 and p185 KO6.2). The p185 Cas9, p185 KO2.3, and p185 KO6.2 cells grown in IL3-free growth medium were treated with imatinib at 0.31, 0.63, 1.25, 2.5, 5, and 10 μM, as indicated, for 48 h, cell viability was determined by trypan blue exclusion assay and represented as average +/- SD of triplicate wells. b. Imatinib-induced cell death in parental and imatinib-tolerant (IMr) p185 Cas9, p185 KO2.3, and p185 KO6.2 cells. The p185 Cas9, p185 KO2.3, and p185 KO6.2 cells are selected without (parental) or with 1.25 μM imatinib (IMr) for six weeks. The cells were then treated with 1.25 μM imatinib for 72 h in IL3-free growth medium. The cell viability was determined by trypan blue exclusion assay and represented as mean +/- SD of triplicate wells. c. Effect of imatinib treatment on the survival of syngeneic mice injected with the control p185Bcr-Abl (p185 Ctrl) and imatinib-tolerant p185Bcr-Abl (p185 IMr) cells. The Balb/C mice were injected through tail vein with 1X106 p185 Ctrl or p185 IMr cells, as indicated. Ten days post-injection the mice were administered intraperitoneally once a day with either saline as control or imatinib (IM, 100 mg/Kg body weight), as indicated, for 5 consecutive days. Survival of the mice were monitored and expressed as the percentage of survival. d. The Abi1 deficient p185Bcr-Abl cells tolerant to imatinib failed to develop leukemia in syngeneic mice. The Balb/C mice were injected through tail vein with 1X106 imatinib-tolerant p185Bcr-Abl control (p185 Ctrl IMr) cells as well as imatinib-tolerant p185 KO2.3 and p185 KO6.2 cells (p185 KO2.3 IMr and p185 KO6.2 IMr). Ten days post-injection the mice injected with p185 Ctrl IMr cells were administered intraperitoneally once a day with imatinib (IM, 100 mg/Kg body weight) for 5 consecutive days. Survival of the mice were monitored and represented as the percentage of survival

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