Research Paper Volume 14, Issue 7 pp 3049—3069

Figure 2. Validation of circGSE1 and differences in organization. (A) Sanger sequencing of the band amplified with divergent primers confirms head-to-tail splicing of circGSE1. (B) Agarose gel electrophoresis validated the expression of circGSE1 in MAECs. CircGSE1 was amplified by divergent primers in cDNA but not gDNA, and GAPDH was used as a negative control. (C, D) In the presence or absence of RNase R, the expression of circGSE1 and linear GSE1 RNA in MAECs was detected by qRT-PCR, and nucleic acid electrophoresis or qRT-PCR was then performed. (E) FISH experiments proved that most circGSE1 is localized in the cytoplasm. Nuclei were stained blue, and cytoplasmic circGSE1 was stained red. (magnification, 400×, scale bar, 20 μm). (F) The circGSE1 expression levels in different mouse organs were analysed by qRT-PCR, revealing higher expression in muscle, artery and splenic organs. (The data are expressed as the mean ± SD, *P < 0.05, **P < 0.01 versus the negative control).