Fig. 2

CAF-secreted EVs exhibit elevated levels of circBIRC6. (A) Assessment of circBIRC6 expression in the culture medium of CAFs and NAFs via qRT-PCR. (B) Quantification of circBIRC6 expression in Panc-1, MiaPaCa-2, and BxPC-3 cells post-treatment with culture medium from CAFs and NAFs. (C) Analysis of circBIRC6 levels in the CAF culture medium following treatment with RNase or a combination of RNase and Triton X-100. (D) qRT-PCR analysis of circBIRC6 expression in Panc-1 and MiaPaCa-2 cells following treatment with CAF culture medium in the presence of α–amanitin, an RNA transcription inhibitor. (E-F) Characterization of CAF-derived EVs using TEM (E) and NanoSight (F). Scale bars represent 200 nm. (G) Western blot detection of EV-derived protein markers (TSG101 and CD9) in CAF cell lysates and EVs. (H) Comparative analysis of circBIRC6 levels in EVs, soluble fractions, and whole culture medium from CAFs using qRT-PCR (n = 3). (I) Correlation analysis between EV-packaged circBIRC6 expression in CAFs and patient plasma samples. Data are mean ± SD. n.s., not significant. ***p < 0.001