Quantitative Western Blot Analysis

Mouse tissues were dissociated using Bio-Plex Cell Lysis Kit (Bio-Rad, Mississauga, ON, Canada). Western blotting was performed as previously described^{68 (link)}. Antibodies against indicated proteins were: phospho-eIF4E (Ser209, Abcam, 1:1000), eIF4E (BD Biosciences, 1:1000), IκBα (Cell Signaling Technology, 1:1000), and glyceraldehyde 3-phosphate dehydrogenase (GAPDH, coupled to Horseradish Peroxidase, Abcam, 1:5000) or β-actin (Sigma, 1:5000); secondary antibodies were anti-mouse and anti-rabbit (GE Healthcare). Quantification of immunoblots was performed using ImageJ (NIH), and expressed as a ratio (either p-eIF4E/eIF4E or eIF4E/GAPDH or eIF4E/β-actin, IκBα/GAPDH). Western blots experiments were replicated at least two times. Uncropped western blots are provided in Supplementary Figure 9.