Information S1 - Adenoviral Vector Driven by a Minimal Rad51 Promoter Is Selective for p53-Deficient Tumor Cells

Rad51core promoter activity correlates with host cell expression of Rad51 protein. (A) Nuclear extracts were isolated from each cell type and 3 µg of total protein was separated on a 10% polyacrylamide gel, transferred to a nitrocellulose membrane, and probed with a monoclonal antibody for Rad51. The blots were stripped and re-probed for β-tubulin as a loading control. All displayed samples were run on the same blot. (B) Rad51 expression was quantitated by densitometry from the blot shown in panel (A) using QuantityOne software (BioRad); Rad51 expression was then normalized in terms of housekeeping protein levels (β-tubulin). Rad51 expression levels in normal fibroblasts (Wi-38 cells) were defined as 1, and Rad51 expression levels in the other cells were then expressed relative to this. Shown is a scatterplot of endogenous Rad51 protein content for each cell line, versus the level of Rad51core promoter activity in the same cell line. Statistical analysis was performed using a non-parametric test and determined to be significant (Spearman rank correlation coefficient = 0.7, p = 0.04). (PDF)