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Fig. 2 | Molecular Cancer

Fig. 2

From: The circSPON2/miR-331-3p axis regulates PRMT5, an epigenetic regulator of CAMK2N1 transcription and prostate cancer progression

Fig. 2

CAMK2N1 is a transcriptional target of PRMT5 in PCa cells. A Scatter plot of differentially expressed genes. B Volcano plot of differentially expressed genes. C Heatmap analyzed from RNA-seq data (GEO: GSE56757) listed the differentially expressed genes in LNCaP cells treated with negative control (NT-shRNAs) or PRMT5 shRNAs. D The most 10 upregulated genes in LNCaP cells treated with negative control (NT-shRNAs) or PRMT5 shRNAs analyzed from RNA-seq data (GEO: GSE56757). E Relative protein levels of CAMK2N1 were detected by Western blot in negative control (NC) or PRMT5-depleted (shRNAs) PC-3 and DU145 cells. F Relative mRNA levels of CAMK2N1 were detected by qRT-PCR assays in negative control (NC) or PRMT5-depleted (shRNAs) PC-3 and DU145 cells. G Promoter-driven luciferase reporter assays in PC-3 and DU145 cells transfected with pGL3.0 luciferase reporter plasmid containing promoter region of CAMK2N1 (-2000 to -1 bp) and pcDNA3.1-PRMT5, compared with that of cells co-transfected with pGL3.0-promoter (-2000 to -1 bp) and empty vector. **P < 0.01. H Relative enrichment of PRMT5 and its catalytic histone marks H4R3me2s and H3R8me2s on the promoter region of CAMK2N1 gene was evaluated by ChIP-qPCR assays in PC-3 and DU145 cells. P1 (+ 155 ~  + 254), P2 (-192 ~  + 19), P3 (-564 ~ -417), P4 (-1050 ~ -934), P5 (-1581 ~ -1362), P6 (-1866 ~ -1767) and P7 (-2346 ~ -2239). IgG was used as a negative control. I Relative enrichment of PRMT5 and its catalytic histone marks H4R3me2s and H3R8me2s at the selected loci (P3: -564 ~ -417) of CAMK2N1 promoter was evaluated by ChIP-qPCR assays in control (NC) or PRMT5-depleted PC-3 and DU145 cells. IgG was used as a negative control. Data were showed as mean ± SD. **P < 0.01

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