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. 2015 Aug 14;5(8):e339. doi: 10.1038/bcj.2015.67

Figure 4.

Figure 4

Impacts of Twist1 overexpression on phenotypes of clinical patients and AML cell lines. (a) Kaplan–Meier estimates of overall survival (OS) for AML patients receiving standard treatment according to Twist1 gene expression level. Patients with higher Twist1 expression had significantly longer OS than those with lower expression (P=0.033, log-rank test). (b and c) Roles of Twist1 upregulation on the cellular susceptibility to chemotherapeutic compounds and expression levels of downstream proteins in AML cells. After transient transfection with either a pFLAG-Twist1 plasmid or an empty vector for 48 h, the KG1a cells were exposed to various concentrations of cytarabine (b) and daunorubicin (c) for 72 h for viability assay. The numbers of surviving cells were counted using trypan blue, and the survival curves were plotted. Each value represents the mean±s.d. of three independent experiments. (d) Cellular levels of Bmi1, p14ARF and p16INK4A proteins linked to altered expression of Twist1 in KG1a cells. The KG1a cells, expressing low endogenous Twist1, were transiently transfected with either a pFLAG-Twist1 plasmid or an empty vector for 48 h, and the nuclear lysates were subjected to western blotting analysis. Histone 3 was used as a loading control. Representative data from three independent experiments are presented.