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. 2015 Jul 24;9(4):255–264. doi: 10.1080/19336918.2015.1013383

Figure 4.

Figure 4.

Snail-Rac1 signaling is not regulated by the ERK pathway in 22Rv1 cells. (A) Western blot analysis was performed using LNCaP and 22Rv1 Neo and Snail over-expressing cells or C4–2 cells with stable Snail knockdown, to assess the levels of total and phospho-AKT (p-AKT) and total and phospho-ERK (p-ERK). (B) 22Rv1 Snail-overexpressing cells were treated with 20 µM U0126, MEK inhibitor, for various time points and a western blot analysis performed to measure the total and p-ERK and AKT levels, Snail, maspin and Rac1 protein levels. Western blot data for Snail and maspin was quantified using Image J Software from NIH. (C) Rac1 activity assay was performed following treatment of 22Rv1 Snail-overexpressing cells with UO126, using the G-LISA Rac1 activity assay. (D) A migration assay was also performed after 24 hrs treatment of 22Rv1 Snail-overexpressing cells with the U0126. Actin was used as a loading control. Results are reported as mean ± SD from 3 independent experiments. Statistical significance was assessed using GraphPad Prism software by paired Student's t-test compared to 22Rv1 Snail control cells treated with DMSO (##,**P < 0.01, ***P < 0.001).