Figure 7

Using Western blot to analyze the level of phosphorylated Cofilin (p-Cofilin) expressed in NSC34 cells. (A) Western blot analysis. NSC34 cells were transfected with eno2-siRNA to knock down endogenous Eno2, then rescued with different DNA material, as indicated, in the absence (−) or presence (+) of Pgk1 protein. pCS⁺: plasmid with an siRNA insertion; eno2-wb: wild type Eno2 but consisting of nucleotides modified at wobble position (eno2-wb) to avoid being blocked by introducing siRNA; and D419S: mutant Eno2-wb consisting of an aspartic acid mutated into serine at the 419th position of Eno2. The expressional level of p-Cofilin at Ser3 was quantified relative to that of α-Tubulin, which served as an internal control. The number listed below each lane indicates the fold change of the p-Cofilin intensity of each treatment compared to the control group, set as 1. (B) Quantitative and statistical analyses. Data were averaged from three independent experiments and presented as mean ± SD (n = 3). One-way ANOVA, followed by Tukey’s multiple comparison test, was used to perform statistical analysis (** p < 0.005). F statistics were 4 as numerator and 10 as denominator.