Perlecan function under androgen and growth factor limitation. A. Minimal changes in Perlecan mRNA levels in LNCaP-derived cell lines upon serum starvation. RealTime PCR analysis of Perlecan mRNA levels presented as fold increase in Perlecan under normal (black bars) or starved (grey bars) growth conditions. While Perlecan mRNA is decreased in LNCaP, all other cell lines demonstrate no change in Perlecan mRNA levels. B. Top Panel: No change in Perlecan protein levels upon serum starvation. Agarose based western blots from protein extracts derived from exponentially growing or serum starved LNCaP, C4, C4-2, and C4-2B cells. No significant differences are noted in protein levels between the cell lines or under the differing conditions. Bottom Panel: Equivalent amounts of the same samples loaded on traditional SDS-PAGE and probed for GAPDH as a loading control. C. Increases in expression of SHH and Gli-1 mRNA upon serum starvation. RealTime PCR analysis of SHH (black bars) and GLI1 (grey bars) as increased fold change compared to normal growth conditions. Gene expression determined by All Real Time PCR with an n = 9 and normalized to Beta-actin. Error bars indicate standard deviation.