Fig. 8

Effects of cytokines in EVs derived from Thy1-MCs. Volcano plot of cytokines given by Thy1-MCs and MHs (A). The number of cytokines found in EVs secreted from hepatic Thy1-MCs and MHs was assessed using a custom-designed Quantibody rat-specific protein array (QAR-CAA-67) (B). Higher concentrations of CINC-2 and MCP-1 were found in Thy1-MC-EVs than in MH-EVs. CINC-2 and MCP-1 were added to the culture medium 3 h after plating and refreshed every alternate day. Panel C demonstrates the photos of typical SH colonies treated with each cytokine. BrdU was added to the culture medium 24 h before fixation, and immunocytochemistry for BrdU was conducted 7 days after plating. Scale bar, 100 μm. The total number of SH colonies per well (D) , number of cells per colony (E), and percentage of cells with BrdU⁺ nucleus per colony (F) are demonstrated. Asterisk indicates significant differences at p < 0.05