Fig. 2
ARC and p53 in RCC cell lines. a For ARC knockdown by shRNA in clearCa-12 cells, three lentiviral shRNA vectors were tested, and the shRNA construct pGIPZ 1365 demonstrated the strongest ARC knockdown (approximately 90%) as determined by western blot. b Treatment of clearCa-12 cells with 10 μg/ml topotecan modulated gene expression of 4 out of 26 p53-regulated genes in control cells and 2 out of 26 p53-regulated genes in ARC-knockdown cells, as determined using the p53 Signaling Pathway RT2 Profiler PCR Array (Qiagen, Hilden, Germany) after 12 h of topotecan treatment. Direct comparison of topotecan treated ARC knockdown and control clearCa-12 cells showed no significant differences with regard to the modulation of p53-regulated genes. Genes with a fold-increase in gene-expression greater than 4 are depicted as red points, and those with a fold-decrease greater than 4 are depicted as green points. c ARC knockdown did not change the subcellular localisation of p53 in RCCs. Fluorescent immunohistochemistry for p53 in clearCa-12 cells revealed that ARC knockdown did not change the amount of nuclear p53 in ARC knockdown cells compared to control cells transduced with non-silencing shRNA