Fig 1: Expression of CDC20 and DAPK1 and their correlation with CYP1B1 in clinical samples. a Representative immunostaining of CDC20, DAPK1 and CYP1B1 from matched samples including normal tissue, Low CYP1B1 RCC and High RCC tissues. Intracellular CDC20 accumulation was positive in some cancer cells, whereas majority of the normal kidney cells were negative. Strong cytoplasmic staining of DAPK1 was more common in the normal kidney tissues than in RCC samples. b CDC20 expression was significantly higher in RCC than normal kidney tissues. There was a positive correlation between the expression of CDC20 and CYP1B1 in RCC, though it did not reach statistical significance. DAPK1 expression was significantly down-regulated in RCC as compared to normal tissues. The expression level of DAPK1 was also significantly correlated with Low and High CYP1B1 expression in RCC. *, P < 0.05. **, P < 0.01.***, P < 0.001
Fig 2: Verification of cDNA microarray data by RT-PCR and Western blotting. a Among 9 down-regulated genes, 6 genes (DKC1, OCLN, MIK67, LIG4, CDC20, BIRC5) were confirmed by real-time PCR using Taqman probe in Caki-1 cells. Of these 6 genes, CDC20 was down-regulated in both Caki-1 and 769-P cells. *, P < 0.05. **, P < 0.01.***, P < 0.001. b Among 4 up-regulated genes, TP73 and DAPK1 were confirmed by real-time PCR and only DAPK1 was down-regulated in both cell lines. *, P < 0.05. (C). Western immunoblotting analysis of DAPK1, LIG4 and CDC20 in control and CYP1B1 transfected Caki-1 and 769-P cells. A positive correlation was found between protein levels and mRNA expression of DAPK1 and CDC20 in both cell lines. GAPDH was used as a loading control
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