The expression of PIPKIγ was silenced using three different siRNA sequences in DanG cells for 48 hours. With or without 15 minutes EGF (10 ng/mL) stimulation after overnight serum starvation, cells were subjected to immunoblotting using indicated antibodies to examine the activation of ERK1/2 (A), AKT (B), or STAT3 (C). pERK1/2, phosphorylated ERK1/2; ERK, total ERK1/2; pAKT, phosphorylated AKT; AKT, total AKT; p-STAT3, phosphorylated STAT3; STAT3, total STAT3. ImageJ was used to measure the intensity of each band. Intensities of phosphorylated ERK1/2, AKT or STAT3 bands were normalized against ERK1/2, AKT, or STAT3, respectively to represent the relative activation level of these kinases. Results from three independent experiments were statistically analyzed and plotted as panels shown at the right. *, p<0.05; **, p<0.01.