Figure 3. Loss of TGF-ß responsiveness in prostatic stromal cells causes the prostate to become refractory to androgen ablation.

(A) To assess the competency of Tgfbr2^{FloxE2/FloxE2} and Tgfbr2^fspKO prostatic stromal cells for TGF-ß signaling, Smad2 was localized by Alexa Fluor 594 (red) immunofluorescence following TGF-ß treatment. Hoechst nuclear counter stain was used (blue). (B) H&E staining of prostatic glandular structures in tissue recombination grafts of Tgfbr2^{FloxE2/FloxE2} and Tgfbr2^fspKO stromal cells with epithelial organoids that were allografted for five weeks in intact syngenic host male mice (+ Androgen) (n=8), (C) H&E staining of prostatic glandular structures in tissue recombination grafts of Tgfbr2^{FloxE2/FloxE2} and Tgfbr2^fspKO stromal cells with epithelial organoids that were allografted for five weeks and castrated for the last week of grafting in syngenic host male mice (- Androgen) (n=8). “Kd” indicates kidney tissue. TUNEL staining (indicated with black arrowheads) in Tgfbr2^{FloxE2/FloxE2} and Tgfbr2^fspKO tissue recombination grafts from castrated mice (- Androgen) (n=4). Ki67 staining (black arrowheads) indicate proliferative cells of Tgfbr2^{FloxE2/FloxE2} and Tgfbr2^fspKO allografts from castrated mice (n=4). Open arrowheads indicate background staining of likely dead cells and “Kd” indicates kidney tissue.