Figure 3. Validation of the circRNA-sequencing results. (A) PCR products (eight up-regulated, eight down-regulated and six commonly expressed circRNAs) were examined by 2% agarose electrophoresis. (B) The back-spliced junction of the randomly selected 22 circRNAs was confirmed by sanger sequencing. The black vertical line represented the back-spliced junction sites. The green horizontal line indicated the 5' end and the red line indicated the 3' end of the circRNA sequence. (C) The expression levels of eight differentially expressed (DE) circRNAs in a larger cohort (n = 44) were examined by qPCR. *, P < 0.05; **, P < 0.01; ***, P < 0.001.