A. Quantitative assessment of VHL, PTP1B, Src and HIF-2α expression by immunostaining of RCC TMA. Representative staining images from a patient with strong VHL protein expression (top panel) and from a patient with weak VHL expression (bottom panel) are shown. Corresponding markup images of the color deconvolution algorithm with intensity ranges are shown (Red = strong, orange = moderate, yellow = weak, blue = negative immunoreactivity). For HIF-2α, the nuclear immunostaining algorithm was applied. Scale bar is 50 μm.
B. Spearman Rho correlation coefficients among the biomarkers are listed in the boxes. Red indicates positive correlation and blue indicates negative correlation. P values for these correlations are represented as follows: *p<0.05; **p<0.001; ***p<0.0001 (n = 136).
C. Comparison between Src and VHL protein expression in the RCC tissue microarray
D. Scatter plot of the VHL and Src scores generated from automated image analysis intensity algorithm. The vertical lines represent 5th percentile and median VHL scores, corresponding to thresholds for negative and weak expression, respectively. The horizontal line represents the median for the Src score, where levels below are considered weak expression and levels above are considered strong expression. The upper right (shaded) quadrant depicts the molecular phenotype of tumors with both strong VHL and strong Src expression.