Plot of sensitivity versus specificity.

<p>At a probability cut-off point of 0.45 both the sensitivity (80%) and specificity (85.3%) for this combination of markers is high, indicating excellent discrimination power of the combination.</p

Plot of sensitivity versus specificity.

<p>Area under the ROC curves calculated for combination of FLIP, Sp1, Sp3, Gleason score, and their interactions gives a value of 0.93 indicating excellent discrimination between non-recurrent and recurrent cases.</p

Box plots showing significant differences in mean total score for IHC of Sp1, Sp3, and FLIP between recurrent and non-recurrent cases as determined by Wilcoxon rank-sum test.

<p>Box plots showing significant differences in mean total score for IHC of Sp1, Sp3, and FLIP between recurrent and non-recurrent cases as determined by Wilcoxon rank-sum test.</p

Plot of sensitivity versus specificity.

<p>Area under the ROC curves calculated for FLIP (0.71), Sp1 (0.66), Sp3 (0.68), and Gleason (0.76) show various degrees of discrimination as predictors of recurrence. An area under the ROC curve of 0.8 to 1.0 is considered to be very good to excellent discrimination, whereas 0.5 indicates no discrimination.</p

A. H&E staining and IHC analysis of expression of FLIP, Sp1, and Sp3 in a representative sample of non-recurrent PCA [Gleason 7 (3+4)] under low magnification (left) and high magnification (right).

<p>The total score for this sample was 0, 6, and 0 for FLIP, Sp1, and Sp3 respectively. <b>B.</b> H&E and IHC staining of FLIP, Sp1 and Sp3 in a representative sample from a patient with recurrent PCA [(Gleason 9 (4+5)] under low magnification (left) and high magnification (right). The total score for this sample was 7, 8, and 6 for FLIP, Sp1, and Sp3, respectively.</p

A. Predicted probability of recurrence when Gleason is low grade 5–7(3+4) for different levels of Sp1 (0, 3, and 6) and Sp3 (0–6) as a function of FLIP (0–8) interaction.

<p>Cases above the cut-off point of 0.45 (dashed line) are predicted to recur. The interaction of FLIP and Sp3 is shown as solid color lines on the X-axis. <b>B.</b> Predicted probability of recurrence when Gleason is high grade 7 (4+3) for different levels of Sp1 (0, 3, and 6) and Sp3 (0–6) as a function of FLIP (0–8) interaction. Cases above the cut-off point of 0.45 (dashed line) are predicted to recur. The interaction of FLIP and Sp3 is shown as solid color lines on the X-axis.</p

Univariate analysis of odds ratios for the markers Sp1, FLIP, Sp3, and Gleason score (A) and multivariate analysis showing the main effects and their ability to predict recurrence in combination (B).

<p>Univariate analysis of odds ratios for the markers Sp1, FLIP, Sp3, and Gleason score (A) and multivariate analysis showing the main effects and their ability to predict recurrence in combination (B).</p

FH mRNA expression is reduced in clear cell kidney cancer.

<p>mRNA levels of <i>FH</i> were determined in a separate set of tumor samples relative to patient matched normal renal parenchyma with real time RT-PCR (p = 0.004). Expression levels were normalized to 18 s rRNA levels prior to comparative analysis.</p

FH expression impacts AKT signaling.

<p>A) <i>VHL</i> null 786-O and A498 cells were transfected with siRNA to FH and scramble control (siNC). Forty-eight hours following transfection, protein lysates were analyzed by immunoblotting for total AKT and ser473 phospho-AKT. B) 786-O cells were transiently transfected with control vector (CV) and vector containing FLAG tagged FH. Forty-eight hours following transfection, protein lysates were analyzed by immunoblotting for the indicated proteins. C) 786-O cells were transfected with the indicated siRNA. Twenty-four hours following transfection, media of the cells was replaced with media containing PI3K inhibitor LY-294002 (6.25 µM). Cells were then harvested 24 hours later and protein lysates were subjected to immunoblot analysis for the indicated proteins.</p

HIF-2α promotes migration in RCC cells.

<p>786-O RCC cells were transfected with scramble control siRNA (siNC) or siRNA to HIF-2α. Western blotting results of whole cell extracts are demonstrated in panel A. B) Wound healing assay was performed in transfected cells. Images were serially taken at the indicated time point following “wound” induction. Results are graphically displayed in panel C. Asterisks (*) indicate statistical significance with p<0.05 relative to scramble control transfected cells.</p