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. Author manuscript; available in PMC: 2020 Apr 28.
Published in final edited form as: Cancer Lett. 2019 Feb 1;448:182–196. doi: 10.1016/j.canlet.2019.01.035

Fig. 6.

Fig. 6.

TWIST1 positively regulates LIMK2 protein levels. (A) Overexpression of WT HA-tagged TWIST1 increases LIMK2 levels in C4-2 cells. (B) Histogram shows relative band intensities normalized to the corresponding actin level. Data shown are mean ± SEM of three independent experiments. * and # indicate statistically significant differences with respect to controls for TWIST1 and LIMK2 proteins, respectively. p < 0.05 analyzed by two-way analysis of variance. (C) TWIST1 ablation depletes LIMK2 in C4-2 cells. (D) Histogram shows relative band intensities normalized to the corresponding actin level. Data shown as mean ± SEM of three independent experiment. (E) TWIST1 inhibits LIMK2 degradation. TWIST1-C4-2 and C4-2 cells were treated with cycloheximide (CHX) for 2 h and 4 h, and LIMK2 and TWIST1 levels analyzed. (F) Graphical representation of TWIST1 degradation rate. (G) Graphical representation of LIMK2 degradation rate. (H) TWIST1 stabilizes LIMK2 by inhibiting its ubiquitylation. Each experiment was performed at least three independent times. Representative data are shown.