MSMB and NCOA4 Implicated in Prostate Cancer Risk Figure 1. RNA expression of MSMB and NCOA4 in normal and tumor prostate tissue by rs10994994 genotype. A. Chromosome 10q11 with isoforms of MSMB and NCOA4 (Ensembl build 52). Primers for competitive PCR were designed to cross exon-exon boundaries depicted by colored lines 1–2 (MSMB) and 1–5 (NCOA4). B. Expression in histologically normal prostate tissue (n = 84). Each point represents absolute RNA expression for one individual, normalized to three housekeeping genes. The top and bottom of the boxes within each graph represent the upper and lower quartiles for expression at each genotype. The band inside each box marks the median value. P-value for each graph denotes the significance for association between expression and genotype. C. Expression in prostate tumor tissue in the Dana-Farber Cancer Institute series (n = 61). doi:10.1371/journal.pgen.1001204.g001 Figure 2. Suppressing MSMB or overexpressing NCOA4 is associated with increased anchorage-independent growth of prostate epithelial cells. A. Effects of suppressing MSMB with three independent shRNAs in LHSAR cells (p-values 0.0001; M6, 0.0023; M8, and 0.0001; M9). The increase in anchorage-independent growth inversely correlates with the degree of MSMB suppression (Figure S4). B. Anchorage-independent growth of LHSAR cells overexpressing NCOA4 and a control vector (p = 0.0074). doi:10.1371/journal.pgen.1001204.g002 PLoS Genetics | www.plosgenetics.org 3 November 2010 | Volume 6 | Issue 11 | e1001204