Figure 3

Effects of EGF on VGSC mRNA and protein expression in PC-3M cells. (A) Real-time PCR measurements of Nav1.7 mRNA expression. Data are plotted as fold-differences relative to the control (Cont) level (dotted line). All treatments were for 24 hours. Treatments were as follows: EGF1 (50 ng/ml), EGF2 (100 ng/ml), AG1478 (100 nM), and EGF1/2+AG1478. EGF significantly enhanced mRNA expression, and this was suppressed by co-treatment with AG1478 (100 nM). (B) A typical Western blot of effects on total VGSC protein expression (50 μg of protein per lane) over 24 h, with pan-VGSCα and anti-actinin antibodies. Arrows indicate molecular weights of ~220 kDa (for VGSCα) and ~100 kDa (for actinin). The lanes denote the following: 1) Control (Cont.; cells in 0.5 % FBS); (2) EGF1 (50 ng/ml); (3) EGF2 (100 ng/ml); (4) AG1478 (100 nM); and (5) EGF1 + AG1478. (C) Quantification and averaging of the data shown in (B) with the corresponding conditions (1–5). VGSC protein expression wasquantified from band optical density. Each histobar (in A and C)denotes mean ± standard error (n = 4–6).