Immunohistochemical staining and Western blot analysis of MCP-1 expression.

<p>(<b>A</b>) Normal control group at 14 days after saline solution injection: Light staining of MCP-1 was seen in the renal tissue and a few inflammatory cells and fibrosis were seen in tubulointerstitial. (<b>B</b>) UUO group at 14 days after saline solution injection: The expression of MCP-1 significant increased in the UUO group at 14 days after surgery. (<b>C</b>) hAFSC group at 14 days after hAFSCs injection: The expression of MCP-1 reduced after hAFSCs transplantation. (bar, 50 µm). (<b>D</b>) The total IOD of MCP-1 in each group: The total IOD in each group reflected the level of the MCP-1 protein. The expression of MCP-1 in the UUO group and hAFSC group at day 1 after surgery was not obvious. There was no significant difference in the expression of MCP-1 among the UUO, hAFSC, and control groups at 1 day after surgery. However, the expression of MCP-1 significantly increased in the UUO group and hAFSC group at 3, 7 and 14 days after surgery compared to the control group, respectively. hAFSCs transplantation significantly reduced the tubuloepithelial cell expression of MCP-1 compared to the UUO group. (<b>E, F</b>) Western blot analysis of MCP-1 protein expression. Western blot analysis showed that MCP-1 protein levels began to increase at day 3, and remained elevated at day 14; These results were consistent with the immunohistochemistry findings. hAFSCs transplantation reduced MCP-1 expression associated with UUO(<i>P</i><0.05). Con, the normal control group at day 14; U1, U3, U7 and U14, the UUO group at days 1, 3, 7 and 14, respectively; hA1, hA3, hA7 and hA14, the hAFSC group at days 1, 3, 7 and 14, respectively. Values are presented as the mean±SD. *p<0.05 vs. control group; #p<0.05 vs. UUO group. Values are presented as the mean±SEM. *p<0.05 vs. control group; #p<0.05 vs. UUO group. Values are presented as the mean±SEM. *p<0.05 vs. control group; #p<0.05 vs. UUO group.