Fig. 1: A prostate specific Pten sensitised piggyBac transposon screen.
From: Identification of genes that promote PI3K pathway activation and prostate tumour formation
A a schematic of the transgenes used in this study. The piggyBac (PB) ATP1 transposon, containing PB and Sleeping Beauty (SB) inverted terminal repeats, a CAGGS cytomegalovirus enhancer and chicken beta-actin promoter element, splice acceptor and splice donors (CPbAct SA: Carp β-actin splice acceptor, En2SA: Engrailed-2 exon-2 splice acceptor and SD: Foxf2 exon-1 splice donor) and polyadenylation signals (SV40 pA: bidirectional SV40 polyadenylation signal). A R26PB allele containing the transposase with a loxP flanked stop cassette inserted into the Rosa26 locus. A PBCre4 transgene to drive Cre expression within epithelial cells of the adult prostate. A conditional allele of Pten used in a heterozygous state. Adapted from Rad et al. [7]. B a schematic showing the genetics of the PBCre4; Ptenfl/+; R26PB; ATP1 (PPPA) animals with all four transgenes. C a prostate lesion identified in a Pten heterozygous sensitised PB animal. Left panels; brightfield image of a PPPA prostate indicating a dense focal lesion (red arrow) next to the urethra (white dots). H & E stain of a section of the prostate lesion (black arrowhead indicates an atypical cell) and a control with no lesion. D the number of lesions microscopically identified in each lobe of prostates from PPPA animals. E the number of lesions identified in each animal. The mean is indicated with a horizontal black line. F a schematic of the experimental pipeline. G IHC analysis on sections of two lesions identified in PPPA prostates. Lesion 1 has retained PTEN and has low levels of p-Akt. Lesion 2 has lost PTEN and has high levels of p-Akt, with p-Akt positive cells invading the stroma (black arrows). PTEN and p-Akt staining in a control prostate is shown. All animals are 3 months old.
