Fig. 1

Identification of circPCNXL2 and circPCNXL2 is upregulated in ICC. (a) The volcano plot of differentially expressed circRNAs. (b) Heatmap of the differentially expressed circRNAs in seven ICC tissues and paired normal tissues by circRNA sequencing. (c) Relative mRNA levels of circRCNXL2 in 76 paired ICC and normal tissues by qRT-PCR. (d) The Kaplan-Meier overall survival curve of circPCNXL2 in 76 ICC patients. Patients were stratified by the median expression of circPCNXL2. (e) Relative mRNA levels of circPCNXL2 in BEC, QBC939, HCCC-9810, HuCCT1 and RBE cell lines. (f) Schematic illustration of the genomic location and back splicing of circPCNXL2. Specific divergent primers were designed targeting the back-splice junction and qRT-PCR products were validated by Sanger sequencing. (g) The circular characteristic of circPCNXL2 was detected by qRT-PCR and agarose gel electrophoresis using divergent and convergent primers in cDNA and gDNA. (h-j) The stability of circPCXNL2 and PCNXL2 was evaluated after Actinomycin D or RNase treatment. (k) Nuclear-cytoplasmic fractionation assay identified the localization of circPCNXL2. (l) The subcellular location of circPCNXL2 in ICC cells was investigated by FISH assay, scale bar = 100 μm. *P < 0.05, **P < 0.01, ***P < 0.001