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. 2007 Feb 14;104(8):2897–2902. doi: 10.1073/pnas.0608213104

Fig. 2.

Fig. 2.

PLY treatment induces actin remodeling and small GTPase activation. (A) Phalloidin staining of SH-SY5Y cells after 0.1 μg/ml PLY treatment revealed filopodia, lamellipodia, and stress fiber formation. (Scale bars, 10 μm.) (B) Pull-down analysis of active RhoA, Rac, and Cdc42, visualized by using Western blotting (Left), showed increased RhoA and Rac activation 4, 8, and 16 min (4′, 8′, and 16′) after treatment with 0.1 μg/ml PLY vs. mock-treated controls (Ctrl or 0′). (Right) Bars represent densitometric analysis of five independent experiments. All values represent means ± SEM. ∗∗∗, P < 0.001, repeated measures ANOVA with Bonferroni post hoc test. (C) The specific Rac1 inhibitor NSC23766 (100 μM) inhibited the formation of lamellipodia 8 min after 0.1 μg/ml PLY treatment. All values represent means ± SEM. n = 100 cells for the lamellipodia perimeter length experiments; n = 4 experiments for the relative lamellipodia density evaluation. ∗∗, P < 0.01; ∗∗∗, P < 0.001, one-way ANOVA with Bonferroni post hoc test. (D) The specific ROCK inhibitor Y27632 (10 μM) reverted the increased stress fiber formation 20 min after 0.1 μg/ml PLY treatment. All values represent means ± SEM of five independent experiments. ∗∗∗, P < 0.001, one-way ANOVA with Bonferroni post hoc test.