Fig 2: RASAL2 regulates PCa cell proliferation via PI3K/AKT/cyclin D1 signalling.A Western blotting analysis of AKT, p-AKT and cyclin D1 expression levels in established cell lines. B Western blotting analysis of AKT, p-AKT and cyclin D1 expression in the LNCaP cell line treated with an AKT inhibitor. C Flow cytometry analysis of AKT inhibitor-treated LNCaP cell lines. D Western blotting analysis of cyclin D1 expression levels in the siCCND1-treated LNCaP cell line. Cell viability (E), colony formation (F), cell cycle (G) and cell proliferation (H) assays in siCCND1-treated LNCaP cell lines.

Fig 3: Effect of RASAL2 on the PI3K/AKT pathway and cyclin D1 (CCND1) expression.A, B GSEA for RNA sequencing of PCa sublines with RASAL2 knockdown. C, D Quantitative real-time RT–PCR and Western blotting analysis of CCND1 mRNA expression in established cell lines.

Fig 4: Correlation of RASAL2 and cyclin D1 in clinical specimens.A Immunohistochemical analysis of the expression levels and correlation of RASAL2 and cyclin D1 in clinical samples of PCa (n = 19). B Molecular correlation analysis of RASAL2 and cyclin D1 (CCND1) from the GEO database (GSE29079, GSE8511 and GSE62293). C Correlation analysis of RASAL2 and cyclin D1 (CCND1) from TCGA database.

Fig 5: Expression of RASAL2 in PCa and normal prostate tissues.A Immunohistochemical staining of RASAL2 in prostate cancer, adjacent nontumor tissue and metastatic lymph nodes (n = 19). The scale bar is 100 µm. B RASAL2 mRNA expression in normal prostate tissues and PCa tissues from the GEO database (GSE29079 and GSE68555). C RASAL2 mRNA expression in normal prostate tissues and PCa tissues from Oncomine databases (Tomlin’s and Wallace’s). D Clinical correlation of RASAL2 with PCa T-stage and Gleason score from TCGA database. E Prognostic data for high and low expression of RASAL2 derived from the TCGA database.