Figure 9

Knockdown of SLIT2 enhances the phosphorylation levels of PAKs, RAFs and ERK1/2. The granulosa cells were transfected with the SLIT2-specific siRNA, scrambled siRNA (negative control) or no siRNA (blank control). (A) An in vitro phosphorylation assay was performed by western blotting. β-actin was used as a loading control. The blots were cropped, and the gels were run under the same experimental conditions. (B,C) The blotting signal intensity was quantified densitometrically after phosphorimaging (shown in A) and normalized for loading by comparison to the signal of β-actin. The signal intensity of the targeted proteins or phosphorylated proteins was expressed as a ratio to the β-actin signal in arbitrary units (n = 5 per mean ± SEM). Five independent experiments were carried out in triplicate. The results are representative of at least three independent experiments. The statistical significance is marked with different superscript symbols **P < 0.01, *P < 0.05.