FIGURE 1. High saturated fat diet promotes the synergy of Src(WT)+AR induced prostate tumorigenesis.

(A) Schematic of the in vivo prostate regeneration assay. Prostate tissues were isolated from C57BL/6J mice and were dissociated into single cells. The isolated primary prostate epithelial cells were transduced with Src(WT) and AR by lentiviral infection. The bicistronic lentiviral vector for co-overexpression of Src(WT) and AR was created as shown in the scheme. The transduced prostate epithelial cells were mixed with urogenital mesenchymal cells (UGSM). The cell mixture was implanted under the renal capsule of SCID mice. The SCID mice carrying the grafts were fed a 10%, 45%, or 60% fat diet for 12 weeks (n=4 per group). The dietary components including the amount and type of fatty acids are listed in Supplemental Table 1 and 2. (B) Co-overexpression of Src(WT) and AR in lentiviral vector. 293T cells were transduced with AR, Src(WT), Src(Y529F), or AR+Src(WT) (MOI =1, 10, or 100) by lentiviral infection. Expression levels of AR, Src, and Erk were examined by immunoblotting. (C) Representative images of H&E staining of liver and white adipose tissue derived from the host SCID mice on 10%, 45%, or 60% fat diets after 12 weeks in (A). High fat diets (45% and 60% fat diets) increase the size of white adipose cells and lipid deposits in the liver cells. (D) The regenerated tissues from (A) were subjected to H&E and IHC staining. Representative H&E (a-c) and IHC staining of total Src (d-f), AR (g-i), and co-staining of CK5 (basal cell marker)/ CK8 (luminal cell marker)/DAPI (j-l) in tumors are displayed (scale bar, 100 μm). Yellow asterisks indicate the regenerated tubules or tissue transduced with Src(WT)+AR, based on elevated levels of Src and/or AR in IHC. Red arrows indicate the regenerated tissues being transduced with Src(WT)+AR and yellow arrows indicate the neighboring control tubule not being transduced.