Figure 4.
Silencing of endogenous PLK1 is affinity dependent. A431 cells were transfected with p19-E18 or p19-E18N15K,G16R that were loaded with either PLK1 siRNA or negative control (NEG) siRNA. The concentration of C225.2/PFOT490A,L491V was fixed at 5 nM. After 24 h, expression levels of PLK1 (relative to β-actin) were quantified by qPCR for mRNA (A) or by western blot for protein (B). A representative blot is shown below. (C) Cell viability at 48 h after transfection was measured using the WST-1 reagent. All measurements were normalized to that of control cells treated with C225.2/PFOT490A,L491V only. Shown are the averages of three independent experiments ± S.E.M. for each panel. *P < 0.05, ***P < 0.001, ****P < 0.0001 determined by two-way ANOVA with Dunnett's post- test.