Fig. 3.

YBX1 protein binds hY4F to regulate its sorting into lung cancer EVs. A Schematic representation of the biotin-labelled hY4F RNA pulldown assay. B Representative MS/MS data of specific peptides of YBX1 identified in this paper. C) Western blot analysis of hY4F-binding proteins enriched by the RNA pulldown assay. D Western blot analysis of YBX1 and Ro60 in IMR-90 and A549 cells. E Western blot analysis of siRNA-mediated YBX1 knockdown of A549 cells after transfection for 24 h. And #1, #2, and #3 means different siRNAs targeting YBX1. F Quantitative PCR results of hY4F in EVs and cell lysates from A549 transfected with siRNA (#1) targeting YBX1 for 48 h. G Quantitative PCR results of hY4F in EVs and cell lysates from wild type and YBX1 knockout (KO) A549 cells. H Quantitative PCR data of hY4F in EVs from A549-YBX1 KO cells overexpressing YBX1 or an empty vector. I Quantitative data of wild type and YBX1-KO A549 EVs using the surface protein BCA assay. Data from three independent experiments are shown as the mean ± SD (error bars). *P < 0.05, **P < 0.01, ***P < 0.001. ns: no significant difference (Student’s t-test). NC: negative control