fig5

Figure 5. CircGNB1 interaction with CDA1. (A) AGO2 RIP assay results; (B) Expression analysis of linear GNB1 in cells with circGNB1 knockdown; (C) RNA pull-down and LC-MS/MS analysis workflow; (D) Identified proteins enriched with biotinylated probes; (E and F) Validation of interaction between CDA1 and circGNB1 by RNA pull-down and RIP assays; (G) Colocalization visualized by FISH and immunofluorescence (Scale bar = 5 μm); (H and I) qRT-PCR and western blot analyses of CDA1 expression in circGNB1-depleted cells. ^**P < 0.01; ^***P < 0.001. AGO2: Argonaute 2; RIP: RNA immunoprecipitation; LC-MS/MS: liquid chromatography-tandem mass spectrometry; FISH: fluorescence in situ hybridization; qRT-PCR: quantitative reverse transcription polymerase chain reaction; CDA1: cell division autoantigen 1; DAPI: 4’,6-diamidino-2-phenylindole; circGNB1: circular RNA derived from the GNB1 gene; sh-NC: short hairpin RNA negative control; GAPDH: glyceraldehyde-3-phosphate dehydrogenase.