Fig. 4

hnRNPA2B1 plays a role in the transfer of CRC cell-derived exosomal miR-934 to macrophages a qPCR and western blot analysis of hnRNPA2B1 expression in HCT-8 and LoVo cells after hnRNPA2B1 knockdown. b qPCR analysis of miR-934 expression in cell lysates and exosomes derived from hnRNPA2B1-knockdown HCT-8 and LoVo cells. c Western blot analysis was performed to determine the association between biotinylated wild-type or mutant miR-934 and hnRNPA2B1 expression in samples derived by miRNA pull-downs performed with nuclear, cytoplasmic, or exosomal extracts from the HCT-8 and LoVo cell lines. Biotinylated poly(G) served as the negative control. d RIP assays with anti-hnRNPA2B1 antibody were performed on HCT-8/LoVo-CM and HCT-8/LoVo-derived exosomes. IgG served as the negative control. qPCR was then used to analyze the expression levels of miR-934 in the immunoprecipitated samples, and the expression levels of miR-934 were expressed as percentages with respect to the input sample (% input) in HCT-8 and LoVo cell lines. e HCT-8 cells and LoVo cells cotransfected with 100 nM Cy3-labeled miR-934 and sh-hnRNPA2B1 or negative control and then cocultured with PMA-pretreated THP-1 cells for 12 h. An immunofluorescence assay was performed to analyze the effects of hnRNPA2B1 knockdown on the transfer of exosomal (Cy3-labeled) miR-934 from HCT-8 and LoVo cells to macrophages (*p < 0.05; **p < 0.01; ***p < 0.001)