Fig. 2

CircARID1A interacts with IGF2BP3 protein in GC cells. A Genomic locus of circARID1A in ARID1A. Sanger sequence of PCR products of circARID1A amplified in SGC7901 cells was shown. B Existence of circARID1A in cDNA and genomic DNA (gDNA) in GC cells. C Stability of circARID1A and ARID1A in SGC7901 and BGC823 cells treated with actinomycin D. D Expression of circARID1A and ARID1A for total RNA treated with RNase R. E Expression of circARID1A, ARID1A, GAPDH and snoU6 in nuclear and cytoplasmic fractions of SGC7901 and BGC823 cells. F CircARID1A location in SGC7901 and BGC823 cells evaluated by RNA-FISH. G RIP analysis of circARID1A enrichment pull-downs by GFP in SGC7901 cells overexpressing EGFP-tagged IGF2BP3. H-I RIP analyses of circARID1A enrichment pull-downs by IGF2BP3 in SGC7901 cells following (H) IGF2BP3 and (I) circARID1A knockdowns. J Enrichment efficiency of biotin tagged circARID1A probes assessed by RNA pull-downs. K Western blot validation of interaction of circARID1A and IGF2BP3 by RNA pull-down in SGC7901 cells. L RNA-FISH combined with IF demonstrating colocalization of circARID1A and IGF2BP3 in SGC7901 and BGC823 cells. M Schematic of IGF2BP3 binding domains. N RIP analysis of circARID1A enrichment pull-downs by GFP in SGC7901 cells transfected with full-length of IGF2BP3 or truncations. The P-values were calculated using the two-tailed Student’s t test. *P < 0.05