Fig. 1

Identification and Validation of a Novel Circular RNA (circMRPS35) in Human Gastric Cancer Tissues. a Heat map of all differentially expressed circRNAs. Red represents upregulated circRNAs, and green represents downregulated circRNAs in gastric cancer tissues compared to the paracancer tissues. b The back-spliced reads distribution of identified circRNAs in three paired gastric cancer tissues and adjacent normal tissues. c The length distribution of the identified exonic circRNAs. d Genomic origin of the above circRNAs. e Heat map of the significantly differentially expressed circRNAs from a. f The differentially expressed circRNAs validated in 30 pairs of gastric cancer tissues and adjacent normal tissues by qRT-PCR assay. g Schematic diagram of circMRPS35 and the design of PCR primers. h PCR validation of the circMRPS35 amplified by divergent primers and convergent primers using the template cDNA and genomic DNA (gDNA) derived from MKN45 cells. β-actin, linear control. i The Sanger sequencing of the back-splice sites of the products from h. The red arrow represents the “head-to-tail” splicing sites of circMRPS35. j qRT-PCR analysis for the circMRPS35 and MRPS35 mRNA using the template cDNA reverse-transcribed by random primers and oligo dT primers. k qRT-PCR assay for the expression of circMRPS35 and MRPS35 mRNA in MKN45 cells treated with RNase R. l qRT-PCR assay for the expression of circMRPS35 and MRPS35 mRNA in MKN45 cells treated with the transcription inhibitor Actinomycin D (2 μg/ml) at the indicated time points. *P < 0.05, **P < 0.01, ***P < 0.001