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. 2009 Nov 23;4(11):e7953. doi: 10.1371/journal.pone.0007953

Figure 6. The role of Erk1/2 and Smad2 in cystatin C regulated invasion of PC-3 cells.

Figure 6

A). Immunoblot analysis of P-Smad2 in PC-3 cells after silencing of Smad2. B–C). Invasion assay in PC-3 cells after silencing of Smad2 simultaneously with knockdown of cystatin C. The representative pictures are shown in B and quantitative results of 3 independent experiments are presented in C. *p<0.05 (Student T-test). D). Immunoblot with antibody against phosphorylated (Ser383)-Elk1 and cystatin C in the lysates from PC3 cells transfected transiently with siRNA cystatin C and control siRNA and co-treated with Erk2 inhibitor (25 µM). Note that Erk2 inhibitor blocks downstream phosphorylation of Elk1 a target of Erk2 in cells with knockdown of cystatin C. Data are representative for 2 experiments. E–F). Invasion assay in PC-3 cells after concomitant silencing of cystatin C and inhibition of Erk2 with selective inhibitor. The representative pictures are shown in D and quantitative results of 2 independent experiments are presented in E. *p<0.05, #p<0.01 (Student T-test).