Analysis of FOXA1 and uroplakin expression in vitro and in human bladder tissue.

<p>A: A panel of commonly used urothelial cell lines was screened via traditional RT-PCR for the presence of FOXA1, FOXA2, and FOXA3 transcripts, as well as members of the uroplakin (UPK) family. HepG2 cells, which express each member of the FOXA subfamily, were used as positive controls (data not shown). RT4 cells exhibited robust expression of FOXA1 and as previously reported, UPK family members <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0036669#pone.0036669-Olsburgh1" target="_blank">[20]</a>. FOXA2 expression was detected in T24 cells, but was not correlated with UPK family member expression. FOXA3 was not detected in any tested cell line (data not shown). B:Q-RT-PCR analysis shows FOXA1 expression in the SCaBER cell line derived from a primary SCC and in T24 is significantly lower compared to RT4. C∶Decreased FOXA1 expression is associated with decreased UPK expression in human tissue. Archival normal adjacent tissue (top panel) and muscle invasive bladder tumor was immunostained with a pan-UPK antibody, AUM <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0036669#pone.0036669-Huang1" target="_blank">[24]</a> as well as an antibody directed against FOXA1. Out of 10 samples with FOXA1 positive NAT, and FOXA1 negative associated tumor, 4 tumors exhibited negative AUM staining.