Blockade of TRPM7 Channel Activity and Cell Death by Inhibitors of 5-Lipoxygenase (original) (raw)
Figure 3
The effect of 5-LOX on TRPM7-mediated cell rounding and TRPM7 current density.
(A) A western blot using an antibody against 5-LOX revealed the efficiency of the dsiRNA-mediated knockdown of endogenous 5-LOX in 293-TRPM7 cells. 293-TRPM7 cells were transfected with 10 nM dsiRNAs targeting 5-LOX (dsiLOX5-1 & dsiLOX5-2) and the non-silencing control dsiRNA (dsiCT). Cells were lysed 48 hours after transfection. A western blot of β-actin is shown to demonstrate the equal loading of samples. (B) Transfection of dsiRNAs targeting 5-LOX modestly decreased TRPM7-mediated cell rounding. (C) Quantification of the degree of cell rounding under the conditions depicted in (B). Values are presented as the mean ± standard deviation of three independent experiments. The asterisk indicates a significant difference in cell rounding between 5-LOX knockdown (dsiLOX5-1 & dsiLOX5-2) and control cells (dsiCT), using Student's t test (p<0.05) (D) Knockdown of 5-LOX did not appreciably decrease TRPM7's current density at +100 mV in 293-TRPM7 expressing cells transfected with dsiLOX5-2 compared to cells transfected with the control (dsiCT). _p_ = 0.2869 was compared with control cells (dsiCT) using Student's _t_ test (_p_>0.1).