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. 2015 Jul 22;6(29):28223–28237. doi: 10.18632/oncotarget.4950

Figure 7. BCR-ABL decreases ZNF224 expression via PI3K-Akt signaling pathway.

Figure 7

a. K562 cells were treated for 16h with 1 μM Imatinib, or 30 μM PI3K inhibitor (LY294002), or 20 μM dual PI3K/AKT/mTOR inhibitor (BEZ235), or 50 ng/ml mTOR inhibitor (Rapamycin) or 1 μM mTOR inhibitor (Everolimus) or vehicle only (DMSO) as control. ZNF224 and WT1 mRNA levels were measured by RT-qPCR. Error bars represent standard deviations of three independent experiments. b. Protein extracts were subjected to Western blotting with antibodies against phospho-Akt (Ser473), total Akt, phospho-p70 S6-kinase and total p70 S6-kinase. GAPDH was used as loading control. One representative blot out of two performed is presented. Molecular weights to the left. c. K562 cells were incubated for 16h with 30 μM LY294002 or vehicle only (DMSO) as control, after which 4 μg/ml actinomycin D was added to block de novo mRNA synthesis. Cells were collected at 0, 0.5, 1, 3, 5 and 7h after addition of actinomycin D and RT-qPCR analysis of ZNF224 mRNA levels was performed. Error bars represent standard deviations of three independent experiments.