Figure 4.

Loss of epsins 1 and 2 results in reduced tumor growth and increased survival rate in spontaneous TRAMP tumor model. (a) Generation of Epn1^fl/fl. The diagram shows homologous recombination of the floxed gene-targeting vector at the Epsin 1 (Epn1) locus. Wild type Epn1 allele, top row; targeting construct, second row; targeted Epn1 allele, third row; and Epn1 floxed allele without Neo cassette (Epn1^fl/fl), fourth row. (b) Strategy to generate tamoxifen inducible DKO (iDKO) by crossing Epn1^fl/fl; Epn2^−/− with ER^T2Cre deleter mice. (c) Strategy to generate TRAMP-WT and TRAMP-iDKO mice by crossing TRAMP-tg mice with WT or Epn1^fl/fl; and Epn2^−/−; ER^T2Cre (iDKO) mice. (d) H&E staining of prostate tissue from TRAMP-WT and TRAMP-iDKO mice dissected at 22 weeks. (e) Percentage of prostate tumorigenesis in TRAMP-WT and TRAMP-iDKO mice dissected at 22 weeks. (f) Male urogenital tract of TRAMP-WT and TRAMP-iDKO mice show tumor in prostate of TRAMP-WT but not in TRAMP-iDKO. Black dotted lines indicate tumors. Mice were sacrificed at 30 weeks. (g) Quantification of prostate tumor volume from TRAMP-WT and TRAMP-iDKO mice dissected at 30 weeks. (h) Percentage of male mice surviving in presence and absence of epsins 1 and 2 as a Kaplan-Meier plot. n = 20 in (d)–(g) and n = 10 in (h). *indicates P-value <0.05.