Polyclonal anti erk1 2

The immunoblot analysis was conducted using an enhanced chemiluminescence (ECL) system (Millipore, Billerica, MA, USA), and images were analyzed with the LAS-4000 imaging system (Fujifilm, Tokyo, Japan). Primary antibodies used in this study included polyclonal anti-RBM4 (Santa Cruz Biotechnology, Santa Cruz, CA, USA), monoclonal anti- PTBP2 (Abnova, Taipei, Taiwan), monoclonal anti-PTBP1 (EMD, Millipore), monoclonal anti-Nova1(Abnova), monoclonal anti-Actin (EMD, Millipore), monoclonal anti-FLAG M2 (Sigma-Aldrich, St. Louis, MO, USA), polyclonal anti-ERK1/2, and monoclonal anti-phospho-ERK1/2 (Cell Signaling Technology, Danvers, MA, USA). Intensities of the detected signals were determined using TotalLab Quant Software.

The immunoblot analysis was conducted using an enhanced chemiluminescence (ECL) system (Millipore, Billerica, MA, USA), and images were analyzed with the LAS-4000 imaging system (Fujifilm, Tokyo, Japan). Primary antibodies used in this study included polyclonal anti-RBM4 (Santa Cruz Biotechnology, Santa Cruz, CA, USA), monoclonal anti-nPTB (Abnova, Taipei, Taiwan), polyclonal anti-GAPDH (MDBio, Taipei, Taiwan), monoclonal anti-FLAG M2 (Sigma-Aldrich, St. Louis, MO, USA), polyclonal anti-ERK1/2, polyclonal anti-AKT, monoclonal anti-phospho-ERK1/2 and monoclonal anti-phospho-AKT (Cell Signaling Technology, Danvers, MA, USA).

Cell pellets were collected, and protein lysate was prepared. Western blot and densitometric analysis were performed as previously reported (30 (link), 31 (link)). The primary antibodies used were polyclonal anti-p21^CIP1/waf1 (Santa Cruz), polyclonal anti-Insulin Receptor (Santa Cruz), polyclonal anti-ERK1/2, anti-pERK1/2^Y204, anti-pIR^Y1136/1136 (all from Cell Signaling), monoclonal anti-FLAG and anti-Actin (both from Sigma). Appropriate horseradish peroxidase-conjugated secondary antibodies (Amersham Biosciences) were added and protein expression was then detected using the WesternSure Premium Chemiluminescent Substrate. Immunoblot images were acquired by C-Digit (both from Licor).