Fig. 5

Exosomal hsa-circ-0004771 in CRC cell culture media regulates miR-653/ZEB2 axis. A The morphology of exosomes from cell culture media was identified by TEM. Scale bar: 200 nm; magnification: 30000 × . B The protein expression of TSG101 and CD63 (exosome surface markers) measured by WB. C qRT-PCR data showing upregulation of exosomal hsa-circ-0004771 in CRC cell culture media. (** vs NCM460, P < 0.01). D qRT-PCR and WB showing the upregulation of the relative mRNA and protein expression of ZEB2 in CRC cells, respectively. (** vs NCM460, P < 0.01). E Spearman correlation analysis showing the association between exosomal hsa-circ-0004771 expression in CRC cell culture media and miR-653/ZEB2 expression in CRC cells, and between miR-653 and ZEB2 in CRC cells. F–H qRT-PCR data showing decrease in hsa-circ-0004771 in CRC cells and exosomal hsa-circ-0004771 in cell culture media, and an increase of miR-653 in CRC cells after transfection with hsa-circ-0004771 (** vs siNC, P < 0.01). I qRT-PCR and WB data showing decrease in both ZEB2 mRNA and protein expressions after transfection with hsa-circ-0004771d. (** vs siNC, P < 0.01). J Spearman correlation coefficient analysis showing the relationship between exosomal hsa-circ-0004771 expression in cell culture media and miR-653/ZEB2 expression in CRC cells transfected with si-hsa-circ-0004771, and between miR-653 and ZEB2 in CRC cells transfected with si-hsa-circ-0004771. K Transfection of miR-653 mimic into CRC cells decreased the expression of exosomal hsa-circ-0004771 in cell culture media (** vs miR-NC, P < 0.01). L qRT-PCR and WB data showing the decrease in both ZEB1 mRNA and protein expression after transfection of CRC cells with miR-653 mimic (** vs miR-NC, P < 0.01). ZEB2 zinc finger E-box binding homeobox 2, GAPDH glyceraldehyde-3-phosphate dehydrogenase, mRNA messenger RNA, si small interfering, NC negative control