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. 2014 Jan 30;105(3):297–307. doi: 10.1111/cas.12339

Figure 1.

Figure 1

(Next page) Thyrosine kinase inhibitors (TKI) increases levels of DNA methyltransferases (DNMT). (a) MTT assay. K562 cells were plated in 96-well plates and cultured with dasatinib (10 nM) or nilitinib (100 nM). At the indicated time point, their proliferation was measured by MTT assay. Results represent the mean ± SD of three experiments performed in triplicate. (b, c) Real-time RT-PCR. RNA was extracted from K562 cells. cDNA were synthesized and subjected to real-time RT-PCR to measure the levels of the indicated gene. Results represent the mean ± SD of three experiments performed in triplicate. The statistical significance was assessed using a paired t-test. **P < 0.01; *P < 0.05. (d) Western blot analysis. These cells were harvested, and subjected to western blot analysis to monitor the levels of the indicated proteins. Each lane was loaded with 30 μg of whole protein lysate. Band intensities were quantified using ImageJ software (Wayne Rasband, NIH). (e) Expression of miRNA. Expression of miRNA of K562 cells treated with dasatinib or nilotinib (0–12 months) was analyzed using an Mir-X miRNA qRT-PCR SYBR Kit to measure the levels of the indicated gene. Results represent mean ± SD of duplicate cultures. (f) Real-time RT-PCR. RNA was extracted from bone marrow mononuclear cells isolated from Ph+ ALL (n = 1) and CML (n = 7) patients before and after treatment with TKI. cDNA were synthesized and subjected to real-time RT-PCR to measure the levels of the indicated genes. Results represent mean ± SD of duplicate cultures. **P < 0.01; *P < 0.05.