Figure 10

PLCδ4 siRNAs knock down PLCδ4 expression but do not affect the upregulation of EGFR and the activation of Erk1/2. SiRNAs against varaious targets were obtained from Dharmacon Research (Lafayette, CO). See Methods section for sequence specifics and siRNA transfection procedure. Anti-PLCδ4 polyclonal antibodies raised in rabbits immunized with a synthetic peptide WEQQQTMARHLTEI corresponding to amino acids 390–402 of human PLCδ4 were obtained from Covance Research Products (Berkeley, CA). Antibodies against phosphorylated and non-phosphorylated Erk1/2, EGFR, and Lamin A/C were from Cell Signaling Technology (Beverly, MA). Hsp90 antibodies were from Biosource International (Camarillo, CA). β-actin antibodies were from Santa Cruz Biotechnology (Santa Cruz, CA). Western blot analyses were performed using the ECL Plus™ detection reagents from Amersham Biosciences (Piscataway, NJ).