NFAT5 Contributes to Osmolality-Induced MCP-1 Expression in Mesothelial Cells
Figure 1
Osmolality-induced MCP-1 expression. (a) For determination of MCP-1 secretion, confluent Met5A cells were incubated in isosmotic medium (gray column; 300 mosm/kg H2O) or were exposed to hyperosmotic medium (black column; 400 mosm/kg H2O). Medium osmolality was elevated by addition of glucose, NaCl, or mannitol as indicated. At the indicated times, medium samples were collected and the concentration of MCP-1 in the cell culture supernatant determined by ELISA as described in Section 2. Means ± SEM for per point; * versus isosmotic control. (b) For determination of MCP-1 transcription, confluent Met5A cells remained in isosmotic medium (gray column; 300 mosm/kg H2O) or were exposed to hyperosmotic medium (black column; 400 mosm/kg H2O). Medium osmolality was elevated by addition of glucose, NaCl, or mannitol as indicated. At the indicated time points, RNA was extracted from the cells and the abundance of MCP-1 mRNA transcript was determined by qRT-PCR as described in Section 2. Relative MCP-1 mRNA abundance was normalized to that of β-actin to correct for differences in RNA input. Data are means ± SEM for per point; * versus isosmotic control.
