Fig. 2

CRC_sEVs alter the functional properties and morphology of hepatocytes. a CellTox assay showed that treatment for 24 and 48 h with CRC_sEVs did not alter the viability of hepatocytes (RFU: relative fluorescence units). b Gene expression levels of ALBU, CYP3A4, and APOE were measured in THLE-2 cells treated with CRC_sEVs. c ELISA of ALBU released in the conditioned medium of hepatocytes treated with CRC_sEVs for 24 h. In all reported graphs, the asterisks indicate significant differences vs untreated control cells (Ctrl) (*p < 0.05; **p < 0.01; ***p < 0.01). d Fluorescent confocal microscope images showing HNF4 expression and localization in hepatocytes treated for 24 h with CRC_sEVs; e Fluorescent confocal microscope images showing the morphological changes induced by treatment with CRC_sEVs for 24 h; Actin Green (green) was used to stain actin fibers; Hoechst (blue) was used to stain the nuclei; the yellow arrows indicate the CRC_SEV-induced holes in the hepatocyte monolayer. Ctrl untreated control cells