Fig 2: PNP expression was suppressed by si-PNP-1 and si-PNP-2 transfection at both the mRNA and protein levels in PCa cell lines, PC3 and DU145. (A) PNP mRNA expression 72 h after transfection of the si-RNAs. GAPDH expression was used for normalisation. (B) PNP protein expression 72 h after transfection of the si-RNAs. GAPDH was used as a loading control. The expression ratio of PNP/GAPDH was evaluated using ImageJ software (ver. 1.43; http://rsbweb.nih.gov/ij/index.htmL).

Fig 3: Immunohistochemical staining of PNP in PCa (n=60), PIN (n=10), and normal prostatic tissue (n=10) by tissue microarray (left panel, original magnification × 100; right panel, original magnification × 400). (A) Strongly stained tumour lesion (Gleason Score 3+4, pT3N0); (B) weakly stained PIN lesion; and (C) negative staining in hyperplastic tissue without malignancy. Right side of the figure: Quantification of PNP expression. Expression of PNP was upregulated in PCa specimens compared with normal hyperplastic tissue (P=0.0037).

Fig 4: PNP expression was suppressed by miR-1 and miR-133a transfection at both the mRNA and protein levels in PCa cell lines, PC3 and DU145. (A) PNP mRNA expression after 72 h transfection with miR-1 and miR-133a. GAPDH expression was used for normalisation. (B) PNP protein expression after 72 h transfection of miRNAs. GAPDH was used as a loading control. The expression ratio of PNP/GAPDH was evaluated using ImageJ software (ver. 1.43; http://rsbweb.nih.gov/ij/index.htmL).

Fig 5: Effects of PNP knockdown by si-PNP transfection on PCa cell lines, PC3 and DU145. (A) Cell proliferation determined by the XTT assay; (B) cell migration activity determined by the wound-healing assay; and (C) cell invasion activity determined by the Matrigel invasion assay. **P<0.0001. *P<0.01.