SOX5 promotes anchorage-independent growth of MDCK cells. (A) Validation of murine Sox5 enrichment in library-transduced and polyhema-selected MDCK cells. Ethidium bromide staining of PCR reactions performed on water or on cDNAs from GFP- and library-transduced cells, before and after selection, as indicated. (B) Western blot analysis of human Sox-5 protein expression in hSOX5-transduced MDCK cells or in GFP-transduced control cells, as indicated. Anti-β-actin was used as loading control. (C) Growth curve of GFP- and SOX5-transduced MDCK cells on plastic (adherent) and polyhema (suspension). The x-axis shows the time, the y-axis the percent of viable cells number, compared to 100% at day 0. The data represent the mean and standard deviation of triplicate values. (D) Sox5 promotes growth in soft agar. The bar chart indicates the average and standard error of the number of colonies/field above the threshold for the GFP- or SOX5-transduced cells, as indicated (8 fields/sample, p < 0.0002).