Rabbit anti egfr

PCR-based visualization was performed using Duolink II Rabbit/Mouse Red Kit (Olink Bioscience #92101) following manufacturer's instructions. Primary antibodies used were: rabbit anti-EGFR (GeneTex, Irvine, CA) and anti-polycystin-1 (Santa Cruz Biotechnology, Santa Cruz, CA).

The Jak2 specific inhibitor, AG490 (Sigma-Aldrich), was dissolved in ethanol at a concentration of 5 mM and stored at −20 °C. The working concentration of AG490 in culture medium is 5 μM. Cell lysates were collected by using RIPA buffer containing proteinase and phosphatase inhibitors. The proteins were separated by SDS-PAGE (Sigma-Aldrich) and transferred to a PVDF membrane (Millipore). After blocking by 5% non-fat milk in TBST, the membrane was incubated with appropriately diluted primary antibodies. Rabbit anti-Jak2 (Cell Signaling Technology, Danvers, MA, USA), mouse anti-phospho-Jak2 (Tyr1007/1008) (Cell Signaling), rabbit anti-Stat3 (BD Pharmingen, San Diego, CA, USA), rabbit anti-phospho-Stat3 (Ser727) (Cell Signaling), rabbit anti-EGFR (GeneTex, Irvine, CA, USA), rabbit anti-phospho-EGFR (Tyr1068) (GeneTex), rabbit anti-ERK (GeneTex), rabbit anti-phospho-ERK (Thr202/Tyr2004) antibodies (Invitrogen), rabbit anti-Sox2 (GeneTex), rabbit anti-Oct4 (GeneTex), rabbit anti-Nanog (Biorbyt) and mouse anti-beta-actin (Millipore) were used in this research. The enhanced chemilumescent (ECL) western blotting substrate kit (Thermo) was used to stain the membranes and signals were obtained using UVP BioSpectrum Imaging System (Upland, CA, USA).