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. 2014 Feb 6;110(5):1298–1306. doi: 10.1038/bjc.2014.40

Figure 3.

Figure 3

The effect of DJ-1 overexpression and knockdown on KLF17 and ID-1 expression in breast cancer cells. The basal mRNA expression levels of (A) KLF17 and (B) ID-1 were analysed. MDA-MB-231 and MDA-MB-435 cells were transfected with DJ-1-Flag or siDJ-1 for 24–48 h and mRNA and protein expression levels of (C) KLF17 and (D) ID-1 were analysed using real-time qPCR and western blotting analyses. To evaluate the effect of KLF17 on ID-1 mRNA expression, pCMV6-AC-KLF17-GFP or siKLF17 was transfected for 24 h, after which (E) ID-1 expression was analysed by real-time qPCR. The effect of DJ-1 on KLF17-dependent ID-1 expression (F) was confirmed by ChIP assay. MDA-MB-231 cells were transfected with either DJ-1-Flag or siDJ-1; KLF17 binding to the ID-1 promoter at CACCC nucleototide sequence was then investigated using KLF17 antibody. (G) RT–PCR and real-time qPCR analyses were performed. Input HDAC1 and IgG were used as positive and negative controls, respectively. The statistical significances were evaluated using the Student's t-test (*P<0.05, **P<0.01, ***P<0.001). These results are from three independent experiments; each line represents s.d.