Fig. 1.

Human circRNA microarray analysis was used to characterize circGPRC5A in CRC. A Volcano plot of human circRNA microarray demonstrating upregulated or downregulated circRNAs in five CRC and paired normal tissues. B Human circRNA microarray cluster heat map displaying the top 20 upregulated circRNAs in five CRC and paired normal tissues. C Northern blotting and Sanger sequencing were used to characterize circGPRC5A. D–E CircGPRC5A was amplified from the CRC cell line cDNA (HCT8 and HT29), but not from gDNA, using divergent primers. F–G After RNase R treatment, qRT-PCR was used to confirm the expressions of circGPRC5A and GPRC5A mRNA in HCT8 and HT29 cells. H–I Using oligo dT and random 6-mers, we investigated the levels of circGPRC5A and GPRC5A mRNA expressions in HCT8 and HT29 cells. J–K The expressions of circGPRC5A and GPRC5A mRNA were assessed using qRT-PCR in HCT8 and HT29 cells following treatment with actinomycin D at the appropriate timepoint. The results of each assay, which were carried out three times, are displayed as the mean ± SD. *P < 0.05; **P < 0.01; ***P < 0.001