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. Author manuscript; available in PMC: 2010 Sep 1.
Published in final edited form as: Cell Death Differ. 2009 Oct 16;17(3):469–481. doi: 10.1038/cdd.2009.148

Figure 6. Rescue of phenotypes of mouse PPARγ-deficient prostate epithelial cells by re-expression of either PPARγ1 or PPARγ2 isoform or treatment by Rosiglitazone.

Figure 6

(a) Phase-contrast microscopy morphology of control mPrE-γKO-empty vector and mPrE-γKO-PPARγ1 or -PPARγ2 WT cDNA cells, showing a return to a cobblestone morphology following reintroduction of each PPARγ isoform. IF staining confirmed expression of PPARγ protein in the nuclei of mPrE-γKO-γ1WT and mPrE-γKO-γ2WT cells (inset boxes). β-catenin protein was predominantly nuclear in mPrE-γKO whereas in mPrE-γKO-γ1WT and mPrE-γKO-γ2WT cells the protein was found in the cytoplasm and on intercellular membrane interfaces. Caspase-3 was decreased in mPrE-γKO-γ1WT and mPrE-γKO-γ2WT cells compared to mPrE-γKO-EV cells. Scale bar = 50 μm. (b) Western blot analysis demonstrated PPARγ1 protein in mPrE-γKO-γ1WT and PPARγ2 in mPrE-γKO-γ2WT cells. β-catenin and E-cadherin protein levels increased in mPrE-γKO-γ1WT and mPrE-γKO-γ2WT cells compared to mPrE-γKO-EV control cells. (c) mPrE-γKO-EV cells had decreased mitochondria and increased lipid droplets and secondary lysosomes, similar to mPrE-γKO cells. Scale bar = 2 μm. However introduction of the mouse PPARγ1 wild-type cDNA increased mitochondria above wild-type values and decreased the presence of secondary lysosomes and lipid accumulation. Scale bar = 250 nm. Likewise, introduction of the mouse PPARγ2 wild-type cDNA into mPrE-γKO cells restored the level of mitochondria and reduced the instances of secondary lysosomes and lipid droplets. Scale bar = 500 nm. (d) Tissue recombinants made using control (mPrE-pSIR) or mPrE-γ2sh cells with rat UGM. Sections were examined for secretion by PAS. In control recombinants a normal prostatic phenotype with secretion was noted. In contrast in the mPrE-γ2sh containing recombinants a low grade mPIN (arrow) with less secretion into the luminal space and thickened stromal was seen. p63 and AR protein were decreased in the mPrE-γ2sh containing recombinants, but α-SM-actin protein expression was increased compared to tissue recombinants of mPrE-pSIR. Mice carrying tissue recombinants made by mPrE-pSIR and mPrE-γ2sh cells were administered Rosiglitazone chow (0.005% Rosiglitazone) from the time of grafting until sacrifice at three months. Tissue recombinants of mPrE-γ2sh in these mice (designated mPrE-γ2sh+Rosi.) showed secretion, and increased p63 protein in the basal layer and p63 negative-luminal differentiation by IHC staining (arrows). These recombinants showed well-differentiated prostatic glandular structure and a more normal-appearing stroma. Scale bar = 50 μm in the panels.