Fig. 1.

Hsa_circ_0004296 expression was significantly downregulated in PCa and negatively correlated with cancer metastasis. A Heatmap showing differentially expressed circRNAs (208 upregulated and 230 downregulated circRNAs) between five pairs of PCa and metastatic lymph node tissues (Cas-vs-Control: p value< 0.05 &|log2FC| > 1). B qRT-PCR showed that the expression of circ_0004296 in PCa cell lines was significantly downregulated. C qRT-PCR was used to detect the expression level of circ_0004296 in 40 paired PCa tissues and adjacent normal tissues. D qRT-PCR was used to detect the expression of circ_0004296 in urine from 46 patients with benign prostatic hyperplasia, localized PCa, and metastatic PCa. E qRT-PCR was used to detect the expression of circ_0004296 in plasma from 39 patients with nontumor, localized PCa, and metastatic PCa. F Northern blotting showed that circ_0004296 could be detected only in cDNA and not in gDNA. G Schematic map of circ_000429 formation and Sanger sequencing of the circ_0004296 splice junction. H, I RNase-R and actinomycin D treatment assays showed no change in the expression level of circ_0004296, while the expression of ETS1 was significantly decreased. *P < 0.05, **P < 0.01, ***P < 0.001