Fig. 1

Validation and characteristics of circ_0003204 in HAECs. a The expression of circ_0003204 and circ_0004543 in HAECs treated by ox-LDL (50μg/ml) for 24 h. b Schematic illustration showing USP36 exons 16–17 circularization to form circ_0003204 (black arrow). The presence of circ_0003204 was validated by qRT-PCR, followed by Sanger sequencing. The red arrow represents head-to-tail circ_0003204 splicing sites. c QRT-PCR analysis for circ_0003204 expression in HAECs treated with or without RNase R digestion. The relative expression of circ_0003204 and USP36 mRNA were normalised to the values measured for mock treatment. d QRT-PCR analysis of circ_0003204 and USP36 mRNA in HAECs treated with Actinomycin D at the indicated time points. e QRT-PCR analysis of circ_0003204 and USP36 mRNA in the cytoplasm or the nucleus in HAECs. f FISH indicated the localization of circ_0003204 in the cytoplasm. Cytoplasmic circ_0003204 probe was stained with Cy3 for red color. Nuclei were stained with DAPI for blue color. Scale bar = 20 μM. DAPI, 4′,6-diamidino-2-phenylindole; FISH, fluorescence in situ hybridization; qRT-PCR, real-time quantitative PCR. The values are expressed as mean ± SD from three independent experiments, *P < 0.05