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. 2023 Feb 2;114(4):1582–1595. doi: 10.1111/cas.15723

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© 2023 The Authors. Cancer Science published by John Wiley & Sons Australia, Ltd on behalf of Japanese Cancer Association.

This is an open access article under the terms of the http://creativecommons.org/licenses/by-nc-nd/4.0/ License, which permits use and distribution in any medium, provided the original work is properly cited, the use is non‐commercial and no modifications or adaptations are made.

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Circular RNA ARHGAP5 (circARHGAP5) interacted with AUF1 in cervical squamous cell carcinoma. (A) Ago2 RIP for detection of circARHGAP5 in ME‐180 cells. PDP2 acted as a positive control, while U1 was a negative control. (B) Model of RNA pull‐down assay. (C) Enrichment of circARHGAP5 of RNA pull‐down in ME‐180 cells. (D) Silver staining of proteins binding with circARHGAP5. (E) Western blot of RNA pull‐down for detecting the binding protein of top enrichment proteins of mass spectrometry. (F–H) RNA pull‐down and RIP efficiency of AUF1 protein and enrichment of circARHGAP5 in ME‐180 and SiHa cells. Protein efficiency was validated through western blot, RNA enrichment was validated by quantitative RT‐PCR. (I) FISH of circARHGAP5 (red) along with immunofluorescence staining (IF) of AUF1 (green) in ME‐180 and SiHa cells. IF/FISH assay showed that circARHGAP5 was colocated with AUF1 in ME‐180 and SiHa cells. Scale bar, 5 μm. **p < 0.01, ***p < 0.001. IB, immunoblot; M, marker; ns, not significant; scr, scramble.