Fig. 2

Characterization of circDLG1 in gastric cancer. a A scheme illustrating the production of circDLG1. b CircDLG1 was validated by RT–PCR, and its back splicing junction was verified by Sanger sequencing. The existence of circDLG1 was validated in AGS and SGC7901 cell lines by RT–PCR. GAPDH was used as a linear control. c Random hexamer or oligo (dT)18 primers were used in the reverse transcription experiments. The relative RNA levels were analyzed by qRT–PCR and normalized to the value obtained using random hexamer primers. *P < 0.05, Student’s t-test; the experiment was repeated three times. d The relative RNA levels were analyzed by qRT–PCR and normalized to the value detected in the mock group (AGS cells without transfection of RNase R). *P < 0.05, Student’s t-test; the experiment was repeated three times. e The relative RNA levels of circDLG1 and mDLG1 were analyzed by qRT–PCR after treatment with actinomycin D at the indicated time points in SGC7901 cells. *P < 0.05, two-way ANOVA; the experiment was repeated three times. f The expression of circDLG1 and mDLG1 was measured by qRT–PCR in the nuclear and cytoplasmic fractions, respectively. GAPDH and U6 were used as positive controls for the cytoplasm and nucleus, respectively. g RNA FISH for circDLG1 in the gastric cancer cell line SGC7901. Nuclei were stained with DAPI. Scale bar, 20 μm. The data are representative of three independent replicates