Fig. 10

Activation of KCs by Thy1-EVs. Thy1-EVs were administered for 3 h after plating, and the cells were cultured for 48 h. Expression of Cinc-2, Il25, and miR-199a-5p was measured using quantitative reverse transcription polymerase chain reaction. Asterisk indicates significant differences at p < 0.05 (A). SHs were cultured in serum-free media on Matrigel-coated dishes. KC-CM with and without miR-199a-5p mimics and Thy1-EVs were added to SHs 3 h after plating and cultured for 7 days. CM conditioned medium, EV extracellular vesicle, KC Kupffer cell, SHs small hepatocytes. Panel B contains the photos of typical SH colonies treated with each CM. BrdU was added to the culture medium 24 h before fixation, and immunocytochemistry for BrdU was performed 7 days after plating. Scale bar, 100 μm. Total number of SH colonies per well (C), number of cells per colony (D), and percentage of cells with BrdU⁺ nucleus per colony (E) are shown. Asterisk indicates significant differences at p < 0.05