Fig 1: RCC2 is highly expressed in prostate cancer and indicates poor DFS. A RCC2 mRNA expression was compared between 497 prostate cancer samples and 52 normal prostate samples from The Cancer Genome Atlas (TCGA) database. B RCC2 mRNA expression was compared between PCa samples with different regional lymph node metastasis. C The promoter methylation level of RCC2 in PCa and normal samples from TCGA. D RCC2 mRNA expression was compared between prostate cancer samples and normal samples from GEO46602 database. E DFS between PCa patients with high RCC2 expression compared to low RCC2 expression. ***P＜0.001
Fig 2: Hedgehog signaling pathway involves in RCC2-mediated prostate cancer cells. A Volcanic map showed the DEGs in RCC2 knockdown DU145 cells compared to control DU145 cells. B Top 8 signaling were selected in RCC2 knockdown DU145 cells compared to control DU145 cells based on NSE analysis. C GSEA analysis showed the Hh signaling pathway was enriched in RCC2 knockdown DU145 cells compared to control DU145 cells. D, E The protein expression of three genes (GLI1, GLI3, and SUFU) involved in the Hh signaling pathway in DU145 and PC3 cells after (D) RCC2 knockdown or (E) overexpression were detected with western blot. F The protein expression of three CSCs markers (SOX2, Nanog, and CD44) in DU145 and PC3 cells after RCC2 knockdown or overexpression were tested with western blot
Fig 3: RCC2 promotes cell propagation and migration through GLI1 in vitro. A The protein level of RCC2 and Gli1 in DU145 and PC3 cells transfected with Ctrl, RCC2, RCC2 + siGli1, and RCC2 + GANT61, respectively. B CCK-8 assay of cell viability in DU145 and PC3 cells between four groups. C Cell colony generation assay in DU145 and PC3 cells between four groups. D Cell migration ability in DU145 and PC3 cells between four groups. E Cell invasion assay in DU145 and PC3 cells between four groups. F The expression of CSC marker genes, including SOX2, Nanog, CD44. *P < 0.05, **P < 0.01, ***P < 0.001. G. The effect of RCC2 overexpression or knock down on the capacity of oncosphere formation. Representative images. Scale bar, 100 μm
Fig 4: RCC2 drives cell movement and EMT of prostate cancer. A, B Representative images and statistics chart of cell movement in DU145 and PC3 cells transfected with pcDNA-RCC2 and pcDNA-ctrl or siRCC2#1, siRCC2#2, and siCtrl for 24 h were evaluated using the Transwell assay. C, D Representative images and statistics chart of cell invasion of DU145 and PC3 cells. E, F, G, H A wound healing assay was adopted to measure cell migration ability. I, J The expression levels of EMT markers, including Snail, E-cad and Vim, in DU145 and PC3 cells. **P＜0.01, ***P＜0.001
Fig 5: RCC2 drives tumor development by promoting Gli1 in vivo. A Figs of mouse xenograft tumors from the Ctrl, RCC2 overexpression, and RCC2 overexpression + GANT61 groups. B Tumor growth curves in mice among three groups. C Tumor weight of mice among three groups. D The protein expression of RCC2, Gli1, SOX2, Nanog, and CD44 in mice among three groups were tested with western blot. *P＜0.05, **P＜0.01, ***P＜0.001
Supplier Page from Abcam for Anti-RCC2 antibody