Fig. 1. Validation and expression of circ235 in Bca tissues and cells.
A Circ235 expression was observed to be considerably greater in Bca tissues than in adjacent normal tissues. (n = 51). B Correlation between circ235 levels (nhigh = 30, nlow = 43) and overall survival rate for Bca patients. C Circ235 was highly expressed in multiple Bca cell lines in comparison to the control urinary epithelium SV-HUC-1 cell line. D A schematic diagram was constructed to illustrate the generation of circ235, which was derived from CCNY pre-mRNA through exon 7 ~ exon 4 back splicing (hsa_circ_0000235, 315nt), and the splice junction site was verified using Sanger sequencing. E RT-PCR and agarose gel electrophoresis were carried out in T24 cells to validate the existence of circ235. The control was GAPDH. F, G The relative abundance of circ235 and CCNY was analyzed using qRT-PCR in Bca cells exposed to actinomycin D at the specified time point. H, I The relative mRNA expression of circ235 and CCNY was analyzed using RT-qPCR following treatment with RNase R in Bca cells. J, K The relative abundance of circ235 in Bca cells’ cytoplasm and nucleus was determined by qRT-PCR, U6 serving as a control for the nucleus, while GAPDH serving as a control for the cytoplasm. L Circ235 was predominantly sited Bca cells’ cytoplasm based on RNA FISH. Scale bar = 20 μm. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001.