Fig. 6

Pharmacological inhibition of PAK1 diminishes ESCC cell growth, migration and invasion. a KYSE30 and KYSE150 cells were treated with indicated concentration of IPA-3 for 48 h, and then the protein levels of p-PAK1 (T423), PAK1, p-PAK2 (S141), PAK2, Raf1, p-Raf1 (S338), MEK1, p-MEK1 (S298), ERK1/2, p-ERK1/2 (T202/Y204) were detected by Western blotting analysis. b The ESCC cells were exposed to increasing concentrations of IPA-3 for 72 h, the cell viability was measured by MTT assay. c-d KYSE30 and KYSE150 cells were exposed to increasing concentrations of IPA-3 for 48 h. The cells were then washed, and subjected to a drug-free focus formation assay (c) and soft agar assay (d). e KYSE30 and KYSE150 cells were treated with 10 μM of IPA-3 for 0, 24 h and 48 h after scratching. Scale bars: 200 μm. The average gap width was used to evaluate migration. Right, quantitative analysis of the wound relative breadth. The wound breadth was normalized to the initial time point (0 h). Columns and error bars represent mean ± SD (n = 6 per group). *P < 0.05, **P < 0.01, ***P < 0.001, one-way ANOVA with post-hoc intergroup comparison with the Tukey’s test. (F-G) KYSE30 and KYSE150 cells were treated with 10 μM IPA-3 for 48 h, and then underwent transwell migration (f) and Matrigel invasion (g) assays. Left, representative images; Right, quantative analysis from 6 random fields. Scale bars: 50 μm. mean; error bar, SD. ***P < 0.001 by Student’s t test. h IPA-3 diminished the protein levels of MMP-2 and MMP-9 in ESCC cells. KYSE30 and KYSE150 cells were exposed to the various concentrations of IPA-3 for 48 h, the whole cell lysates were subjected to Western blotting analysis