Figure 5. Dependence of hyperosmotic C9 gene expression on NFAT5 activity. A: The NFAT5 inhibitor rottlerin (10 µM) prevented the hyperosmotic (+ 100 mM NaCl) increase in the C9 mRNA level. The mRNA
level was determined with real-time RT–PCR analysis after stimulation of the cells for 2 h. B: Transfection of RPE cells with NFAT5 siRNA (siNFAT5; 10 nM) resulted in a reduction of the NFAT5 mRNA level in RPE cells
cultured in isosmotic medium for 24 h. C: Knocking down the gene expression of NFAT5 with siRNA (siNFAT5; 10 nM) reduced the level of C9 mRNA in cells cultured for
2 h in hyperosmotic (+ 100 mM NaCl) medium compared to nontransfected cells (control). Nontargeted siRNA (siNon; 10 nM) was
used as a negative control. siRNA transfection was done 26 h before hyperosmotic stimulation. The numbers of independent experiments
using cell lines from different donors are indicated in the bars. Significant difference versus unstimulated control: *p<0.05.
Significant difference versus NaCl control: ●p<0.05.