Goat anti mouse or anti rabbit secondary antibody

Cytochrome c oxidase subunit 1 (CcO1), and ATP synthase subunit β (ATP synthase β) were detected in liver homogenates by Western blot using standard techniques as described previously [25 (link), 30 ]. The primary antibodies employed were a mouse anti-mouse polyclonal anti-CcO 1 (1:1000 dilution; Invitrogen, Cergy Pontoise, France), a mouse anti-mouse polyclonal anti-ATP synthase β (1:1000 dilution; Life Technologies, Courtaboeuf, France), and a rabbit anti-rabbit monoclonal anti-β-actin antibody (1:1000 dilution, Sigma Aldrich, Saint-Quentin, Fallavier, France). Horseradish peroxidase-linked goat anti-mouse or anti-rabbit secondary antibody (1:10,000 dilution; containing goat anti-mouse or anti-rabbit secondary antibody, Sigma Aldrich, Saint-Quentin, Fallavier France) was used for CcO1, ATP synthase β, and β-actin, as appropriate. The protein bands corresponding to CcO1, ATP synthase β, and β-actin (by molecular weight identification and specific positive immunoprevalence) were analyzed by means of density profile plots created using NIH ImageJ 1.63 software. Protein band density profiles for CcO1 and ATP synthase β were normalized for protein load by dividing by the matching density profiles for β-actin.

The DRGs and cells were lysed using cell lysis buffer with protease inhibitors (Roche, Penzberg, Germany). Equivalent amounts of proteins were electrophoretically resolved on a denaturing SDS polyacrylamide gel and electro-transferred onto nitrocellulose membranes. The membranes were initially blocked with 5% nonfat dry milk in Tris-buffered saline (TBS) for 2 h and then probed with antibodies against CXCL12, CXCR4, MMP-2, uPA, and NGF. After co-incubation with the primary antibodies at 4°C overnight, the membranes were hybridized with the appropriate goat anti-mouse or anti-rabbit secondary antibody (Sigma-Aldrich) for 1 h at room temperature. Equal protein sample loading was monitored using an anti-β-actin antibody. The probed proteins were detected by using enhanced chemiluminescence (Millipore).

The Western blotting analysis was performed using standard procedures. The following primary antibodies were used in the experiments: anti-PTBP1 antibody (Cell Signaling Technology, Beverly, MA, USA), anti-PTBP2 antibody (Abcam, Cambridge, UK), anti-PTBP3 antibody (Sigma-Aldrich, St. Louis, MO, USA), anti-HNRNPK antibody (Abcam), anti-HNRNPM antibody (Sigma-Aldrich), anti-FUBP3 antibody (Abcam), anti-CPSF2 antibody (Abcam), anti-G3BP2 antibody (Atlas Antibodies), anti-TGFβ1 antibody (Proteintech Group), anti-TGFβ2 antibody (Abcam), anti-p-SMAD2 antibody (Cell Signaling Technology), anti-SMAD2 antibody (Cell Signaling Technology), anti-p-SMAD3 antibody (Cell Signaling Technology), anti-SMAD3 antibody (Cell Signaling Technology), anti-SMAD5 antibody (Abcam), anti-ID2 antibody (Abcam), anti-ZEB1 antibody (Abcam), anti-SNAI antibody (Abcam), anti-E-cadherin antibody (Proteintech Group), anti-Vimentin antibody (Cell Signaling Technology), anti-N-cadherin antibody (Cell Signaling Technology) and anti-GAPDH antibody (Sigma-Aldrich). The blots were incubated with a goat anti-rabbit or anti-mouse secondary antibody (Sigma-Aldrich) and visualized with a commercial ECL kit (Pierce, Rockford, IL).