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. 2009 May 18;83(3):547–557. doi: 10.1093/cvr/cvp153

Figure 5.

Figure 5

TRPM7 silencing enhances eNOS expression via ERK pathway. (A) RT–PCR (left) or immunoblotting (right) showing the effect of TRPM7-siRNA1 on the level of eNOS mRNA and protein in HUVECs. HUVECs were treated with control or TRPM7-siRNA1. 48 h after transfection, RT–PCR was performed using specific primers for TRPM7, eNOS, and GAPDH. For immunoblotting, eNOS protein level was examined 72 h after the transfection by immunoblotting with anti-eNOS antibody. (B) Treatment with ERK inhibitor U0126 reduced expression of eNOS and its enhancement by TRPM7-siRNA1. (C) TRPM7 silencing increased the production of NO as demonstrated by nitrite measurement. This increased production of nitrite was attenuated by 500 µmol/L l-NAME. n = 12–20. *P < 0.05, control vs. TRPM7-siRNA1-treated cells; #P < 0.05, l-NAME-treated vs. untreated cells. (D) Treatment of cells with l-NAME (100 and 1000 µmol/L) did not affect the enhancement of cell growth/proliferation by TRPM7-siRNA1. n = 9–11. **P < 0.01, control vs. TRPM7-siRNA1-treated cells.