Fig. 1

Identification and characteristics of circRHOBTB3 in PDAC cells and tissues. a Schematic illustration of the genomic location and back splicing of circRHOBTB3, with the junction site validated by Sanger sequencing. b PCR and agarose gel electrophoresis experiments confirmed the circular form of circRHOBTB3 using divergent and convergent primers in PDAC cDNA and gDNA samples. c Actinomycin D treatment was applied to evaluate the stability of circRHOBTB3 and RHOBTB3 mRNA in PDAC cells. d circRHOBTB3 exhibited higher stability than RHOBTB3 mRNA in PDAC cells after treatment of RNase R. e FISH experiments showed that circRHOBTB3 predominantly located in cytoplasm in PANC-1 and MiaPaca-2 cells, the circRHOBTB3 probe was labeled with Cy3(red), while nuclei were stained with DAPI. f Nuclear and cytoplasm fractionation assays revealed the subcellular location of circRHOBTB3. g Relative circRHOBTB3 expression in various cell lines of PDAC was determined by qRT-PCR. h Relative circRHOBTB3 expression in PDAC tissues and adjacent normal tissues was measured by qRT-PCR (n = 110). i The Kaplan-Meier overall survival curve of PDAC patients with either high or low circRHOBTB3 expression level in tumor. The samples were imaged at 1000× magnification. Scale bar = 10 μm. All data are presented as the means ± SD of three independent experiments. *p < 0.05, **p < 0.01, ***p < 0.001