Fig. 2

Characterization of circANKRD12 in human cancer cells. a Transcript structure of the ANKRD12 gene. Two predominant circular isoforms (286 bp and 925 bp) were detected sharing the same backsplice junction between Exon8 and Exon2 as represented by the dashed line. The backsplice junction was sequenced for validation. b Divergent primers designed to detect the circANKRD12 backsplice junction amplified cDNA but not genomic DNA (4 pairs of different primers were used to amplify the circular junction). Linear ANKRD12 amplicon was detected in both cDNA and gDNA. M represents 100 bp ladder. c qRT-PCR for relative abundance of circANKRD12 and linear ANKRD12, GAPDH and HIF1 alpha in SKOV3 cells treated with RNase R d Backsplice junction was detected in cDNA synthesized using random hexamer but not oligoDT primed RNA. The data in c is means with error bars representing standard error of the mean (SEM) from three experiments; **P < 0.01 (Student’s t-test)