Figure 1
LXN is highly expressed in non-malignant prostate luminal cells and repressed in high Gleason grade tumours. (A) Western blot data to assess LXN protein expression in the non-malignant human prostate. Data from seven different tissue specimens GAPDH serves as a loading control*. (B) Direct cell sorting from uncultured human prostate tissue into basal, luminal and stromal fractions reveals that LXN is predominantly expressed in prostate luminal cells and 24.25 ± 5.41-fold greater than basal cells (p < 0.01). RT-PCR analysis from four separate tissue samples. (C) mRNA expression analysis of the luminal specific marker PSA confirms the technical success of the FACS approach. The PC3 and LNCaP cell lines were used as basal and luminal cell controls, respectively. (D) Comparison of LXN gene expression following sub-fractionation of primary cultures of basal epithelial cells into stem cells (SC), transit amplifying (TA) and committed basal (CB) cells. Data is representative of four different primary culture samples. (E) Western blot to compare LXN protein expression in normal and malignant prostate tissue. α-SMA and GAPDH were used as indicators of relative stromal abundance and total protein loading respectively*. (F) Scanning densitometry of (E). (G) Gene expression analysis Grasso et al. prostate database reveals that LXN mRNA expression is significantly reduced in metastatic prostate disease compared to normal prostate (p < 0.01). mRNA expression analysis is presented as 2 Δddct (LXN/RPLP0). *Full length, uncropped blots from (A,F) can be viewed in Supplementary Fig. S6.
