1. Volume 66, Issue 5, Pages 956-963 (November 2014)
Carbonic Anhydrase 9 Expression Increases with Vascular Endothelial Growth Factor– Targeted Therapy and Is Predictive of Outcome in Metastatic Clear Cell Renal Cancer 
Grant D. Stewart, Fiach C. O’Mahony, Alexander Laird, Sukaina Rashid, Sarah A. Martin, Lel Eory, Alexander L.R. Lubbock, Jyoti Nanda, Marie O’Donnell, Alan Mackay, Peter Mullen, S. Alan McNeill, Antony C.P. Riddick, Michael Aitchison, Daniel Berney, Axel Bex, Ian M. Overton, David J. Harrison, Thomas Powles 
European Urology 
Volume 66, Issue 5, Pages (November 2014)
DOI: /j.eururo
Copyright © 2014 European Association of Urology Terms and Conditions

2. Fig. 1
Reverse phase protein array (RPPA) differential protein expression results. Box-and-whisker plot showing differential expression analysis of 55 proteins evaluated by RPPA in sunitinib-naïve (light orange) and treated (dark orange) metastatic clear cell renal cell carcinoma samples. *p<0.05, **p<0.01, ***p<0.001.
European Urology  , DOI: ( /j.eururo )
Copyright © 2014 European Association of Urology Terms and Conditions

3. Fig. 2
Results for significantly variable and differentially expressed proteins in test and validation samples from sunitinib-naïve and treated patients. (a) Box-and-whisker plot showing test set reverse phase protein array differential expression results of four key proteins. Medians and interquartile ranges are shown. (b) Box-and-whisker plot showing automated quantitative analysis (AQUA) evaluated protein expression of four key proteins using a validation cohort of 61 sunitinib-treated and 25 pazopanib-treated and untreated paired metastatic clear cell renal cell carcinoma samples. Of the four proteins, carbonic anhydrase 9 (CA9) was the only one with significant differential protein expression (p=0.01). Medians and interquartile ranges are shown. (c) Kaplan-Meier curve showing relationship of CA9 protein expression determined by AQUA in situ analysis (low vs high, as determined using X-tile [15]) in sunitinib/pazopanib-treated patients to overall survival (hazard ratio: 0.260; 95% confidence interval, 0.111–0.608; p=0.001).
European Urology  , DOI: ( /j.eururo )
Copyright © 2014 European Association of Urology Terms and Conditions

4. Fig. 3
Array comparative genomic hybridization and RNA interference carbonic anhydrase 9 (CA9) results. (a) Heat map plotting gains (red) and losses (blue) of CA9. The right-hand bar represents the logarithm of the odds score (−log10) of the adjusted p value (Fisher test), the dashed line represents p=0.05. There were significantly more losses in the treated samples relative to the untreated patient samples (p=0.002). Supplemental Figure 1 provides further description of regional chromosomal changes and Supplemental Figure 2 provides details of the genome-wide changes in gains and losses following sunitinib therapy. (b) RCC11 and (c) CAKI-2 human renal cell carcinoma transfected with either control or CA9 short interfering RNA (siRNA), followed by sunitinib treatment and cell viability analysis 5 d later. Error bars represent standard errors of the mean. To confirm silencing, cell lysates from RCC11 and CAKI-2 siRNA transfected cells were analysed by western blotting using CA9 and β-actin-specific antibodies, as indicated.
VHL=von Hippel-Lindau; Del=deletion; mut=mutant; NC=no change; wt=wild-type.
European Urology  , DOI: ( /j.eururo )
Copyright © 2014 European Association of Urology Terms and Conditions

5. European Urology 2014 66, 956-963DOI: (10.1016/j.eururo.2014.04.007)
Copyright © 2014 European Association of Urology Terms and Conditions

6. European Urology 2014 66, 956-963DOI: (10.1016/j.eururo.2014.04.007)
Copyright © 2014 European Association of Urology Terms and Conditions