A three-protein model for prediction of TGI.

<p>Least Angle Regression (LAR) method identified 3 markers (Ang1, EGF, & Emmprin) sufficient to build a model to predict CVX-060 TGI. Model performance in the training set (A) and testing set (B) of xenograft lines is shown. Statistical significance was not achieved until combining both sets (C). Each dot represents the model predicted TGI% vs. median observed TGI% of a single xenograft line.</p

FACS analysis of apoptosis or necrosis.

<p>Cells were treated with or without 1 µg/mL DBP-maf for 48 hours, propidium iodide and annexin V were added and cells were analyzed using flow cytometry. Results are representative of three experiments.</p

DBP-maf peptide does not inhibit expression of uPAR in PC3M or LNCaPLN3 cells.

<p>LNCaPLN3 (<b>A</b>) and PC3M cells (<b>B</b>) were treated with DBP or DBP-maf (0.001 and 1 µg/mL) and incubated for 24 hours then harvested. RT products (cDNA), identified as uPAR1, 2, and 3, were amplified by real-time quantitative PCR. p<0.05.</p

DBP-maf inhibits tumor cell migration.

<p>LNCaP (<b>A</b>), LNCaPLN3 (<b>B</b>), PC3M (<b>C</b>) or PC3MLN4 (<b>D</b>) cells were added (150,000/well) to the top chamber of a modified Boyden chamber (+/− DBP-maf) with 10% FBS in the bottom chamber. After 6 hours cells were removed that had not migrated and remaining cells were quantitated using an acid phosphatase assay. Results were normalized to control. Experiments were performed a minimum of three times and error is shown as +/− SD. Compared to cell growth without DBP-maf, adding DBP-maf had a statistically significant overall reduction of cell migration at 30% (P = 0.0003) for the combined four tumor cell types. Individual significant reduction rates were found with each of these tumor cell types. Compared to control, significant reduction was seen with DBP-maf at (<b>A</b>) 20% P = 0.0022 (<b>B</b>) 20% P = 0.0029 (<b>C</b>) 10% P = .0045 (<b>D</b>) 30% P = .0094. n = 3.</p

DBP-maf inhibits tumor cell proliferation.

<p>LNCaP (<b>A</b>), LNCaPLN3 (<b>B</b>), PC3M (<b>C</b>) or PC3MLN4 (<b>D</b>) cells were seeded in 24 well dishes overnight, then medium +/− DBP-maf was added with 1% FBS. After 72 hours cells were quantitated using an acid phosphatase assay. Results were normalized to control. Experiments were performed a minimum of three times and error is shown as +/− SD. Compared to control, significant reduction was seen with DBP-maf at (<b>A</b>) 50% P = 0.0001 (<b>B</b>) 50% P = 0.0001 (<b>C</b>) no significant reduction (<b>D</b>) 40% P = .0073. n = 3.</p

DBP-maf inhibits expression of uPAR in PC3M cells.

<p>PC3M, and PC3MLN4 cells were treated with DBP or DBP-maf (0.001 and 1 µg/mL) and incubated for 24 hours then harvested. RT products (cDNA), identified as uPAR1, 2, and 3, were amplified by real-time quantitative PCR. p<0.05.</p

A DBP-maf peptide inhibits tumor cell migration.

<p>LNCaP (<b>A</b>), LNCaPLN3 (<b>B</b>), PC3M (<b>C</b>) or PC3MLN4 (<b>D</b>) cells were added (150,000/well) to the top chamber of a modified Boyden chamber (+/− DBP-maf) with 10% FBS in the bottom chamber. After 6 hours cells were removed that had not migrated and remaining cells were quantitated using an acid phosphatase assay. Results were normalized to control. Experiments were performed a minimum of three times and error is shown as +/− SD. Compared to migration without DBP-maf, adding DBP-maf had a statistically significant reduction of migration at 40% (P<0.0001) for the combination of all four tumor cell types. Individual significant reduction rates were found with each of these tumor cell types. Compared to control, significant reduction was seen with DBP-maf at (<b>A</b>) 30% P = 0.0038 (<b>B</b>) 40% P = 0.0016 (<b>C</b>) 20% P = .0038 (<b>D</b>) 40% P = .0005. n = 3.</p

DBP-maf inhibits protein expression of uPAR.

<p>LNCaP, LNCaPLN3, PC3M, and PC3MLN4 were treated with DBP or DBP-maf and incubated for 24 hours (<b>A</b>) then harvested and immunoblotted using an anti-uPAR antibody. LnCaPLN3 cells at 72 hours (<b>B</b>). p<0.05.</p