Fig. 2

Characters of circRHOBTB3. a The relative circRHOBTB3 or linear RHOBTB3 mRNA abundance detected by qRT-PCR after treated with or without RNase R in three GC tissues. b qRT-PCR for the relative abundance of circRHOBTB3 and RHOBTB3 mRNA in AGS and HGC27 cell lines after treated with RNase R. The amount of circRHOBTB3 and RHOBTB3 mRNA were standardized to the value detected in the mock treatment. c The constitutions of circRHOBTB3 formed by exon6 and exon7 of RHOBTB3 gene illustrated by the schematic diagram. The sequence of back-junction of circRHOBTB3 was validated by sanger sequencing. Red arrow showed the “head-tail” splicing sites of circRHOBTB3. d CircRHOBTB3 verified in three GC tissues and AGS and HGC27 cell lines by RT-PCR. CircRHOBTB3 amplified by divergent in cDNA but not in genomic DNA (gDNA). e qRT-PCR for abundance of circRHOBTB3 and RHOBTB3 mRNA in AGS cell line treated with Actinomycin D at indicated time point. f qRT-PCR value indicating the abundance of circRHOBTB3, U6 and GAPDH in either the cytoplasm or nuclear of AGS and HGC27 cell lines. GAPDH and circRHOBTB3 were normalized to the value measured in cytoplasm. U6 was normalized to the value measured in nuclear. g RNA FISH was conducted to detect circRHOBTB3’s subcellular in HGC27 cell lines. Nuclei was stained with DAPI. 18 s probe was served as positve control. Scale bar, 10 μm