Figure 4

Effects of capsaicin and PI3-K activator on Rac1-GTP expression in B16F10 melanoma cells. (A) Serum-starved B16-F10 cells were treated with capsaicin (0, 50, 100, 150, and 200 µM) for 24 h. (B) Serum-starved B16-F10 cells were treated with capsaicin (100 µM) and/or PI3-K activator (5 ng/ml) for 6 h. Equal amounts of the protein of B16-F10 cell lysates were affinity-precipitated with GTP-PBD bound to glutathione-agarose beads. Precipitated GTP-Rac1 was detected by immunoblotting with anti-Rac1 antibody. Protein expression was quantified by using the Quality-one-4,2,0 image analyzer as described in Materials and Methods. Values are expressed as a percentage of the control, which was defined as 100%. Statistical analysis was performed using a two-tailed Student's t-test for independent variables. Data are plotted as the mean ± SD (^*P < 0.05 vs. control; ^#P < 0.05 vs. PI3-K activator).