Phospho-EGFR-Y1068 monoclonal Ab (mAb) (cat# 2234S), Akt mAb (cat# 9272S), phospho-Akt (Ser473) mAb (cat# 9271S), phospho-PLC-γ1(Ser1248) mAb (cat# 8713), PLC-γ1 pAb (cat# 2822S), glyceraldehyde-3-phosphate dehydrogenase (GAPDH) mAb (cat# 2118), HRP labeled goat anti-rabbit IgG and goat anti-mouse IgG were purchased from Cell Signaling Technology (Danvers, MA, USA). EGFR polyclonal antibody (pAb) (cat# A11577) and β-Tubulin Rabbit pAb (cat# AC015) were provided by ABclonal Technology (Woburn, MA, USA). All the primary antibodies employed for Western blots were at a 1:1000 dilution. The chemical inhibitors used in this study, U73122 (cat# HY-13419), Gefitinib [28 (link)] (cat# HY–50895), and LY294002 [28 (link)] (cat# HY–10108), were provided by MedChemExpress (Monmouth Junction, NJ, USA). U73122 is an inhibitor that can specifically inhibit PLC-γ1 phosphorylation at Ser1248 [19 (link),29 (link)]. Gefitinib is an EGFR inhibitor that can selectively inhibit EGFR tyrosine kinase activity by binding to the adenosine triphosphate (ATP)-binding site [30 (link)]. LY294002 can specifically inhibit Akt phosphorylation by inactivation of its direct activator phosphoinositide 3-kinases (PI3K) [31 ].
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