Figure 2. IL-6 is required for p62’s role in the tumor microenvironment.
(A) Orthotopic injection of TRAMP-C2Re3 cells into the prostates of syngeneic WT and p62 KO mice. Orthotopic tumors were allowed to grow for two months. (B and C) Genitourinary (GU) tract weight (B) and pictures (C) from (A); n = 5 or 6 mice per genotype. Scale bar, 1 cm. (D) “Response to wounding” GSEA plot of enrichment of gene expression in p62 KO orthotopic tumors. (E) α-SMA staining of orthotopic tumors from WT and p62 KO mice. Scale bars, 25 μm (F) RT-PCR of TGFβ in orthotopic tumors from WT and p62 KO mice (n = 3). (G and H) RT-PCR of CAF markers (α-SMA, TGFβ and Vimentin) in FACS-sorted prostate stromal fraction from WT and p62 KO mice (G) and in WT and p62 KO prostate fibroblasts (H); n = 3. (I) TGFβ production in prostate fibroblasts was determined by ELISA. (J) GSEA plots of enrichment of gene expression in p62 KO orthotopic tumors. (K) RT-PCR of inflammatory cytokines in orthotopic tumors of WT and p62 KO mice. n = 5 or 6 animals per group versus WT. (L) IL-6 ELISA in fibroblasts. (M and N) Orthotopic injection of TRAMP-C2Re3 cells into the prostates of mice of different genotypes (n = 5 or 6 mice). GU weights (M) and pictures (N). Scale bar, 1 cm. *p < 0.05, **p < 0.01. ***p < 0.001. Results are presented as mean ± SEM. See also Figure S2.