Fig. 1

Screening and characterizing hsa_circ_0003258 in PCa. A The heat map showing the top 20 down-regulated circRNAs in prostate cancer plasma samples compared with non-tumor plasma samples analyzed by RNA sequencing. B The volcano plot showing the expression profile of circRNAs in prostate cancer plasma samples. C hsa_circ_0003258 expression evaluated by qRT-PCR in plasma samples of prostate cancer patients. D qRT-PCR for the expression of hsa_circ_0003258 in PCa cell lines and tissues. The expression of hsa_circ_0003258 was normalized to GADPH. E Schematic illustration indicating the generation of hsa_circ_0003258 from its host gene, and validation by Sanger sequencing. F PCR assay with divergent and convergent primers showing the amplification of circRNAs from cDNA or genomic DNA (gDNA) of prostate cancer cell lines. G RNA FISH detecting hsa_circ_0003258’s subcellular localization in DU145 and C4–2 cells. Nuclei was stained with DAPI. Scale bar, 10 μm. H QRT-PCR value indicating the abundance of hsa_circ_0003258, U6 and GAPDH in either the cytoplasm or nucleus of DU145 and C4–2 cells. Hsa_circ_0003258 was normalized to GAPDH in the cytoplasm and U6 in the nucleus I QRT-PCR for abundance of hsa_circ_0003258 and ZNF652 mRNA in DU145 cells treated with Actinomycin D at indicated time point. J The relative hsa_circ_0003258 or linear ZNF652 mRNA abundance detected by qRT-PCR after treated with or without RNase R in DU145 and C4–2 cells. ns: Not Significant; ** P < 0.01; *** P < 0.001; **** P < 0.0001