Western Blot Analysis of Protein Lysates

As previously described [29 (link)], protein lysates were separated in 10% polyacrylamide gel, and transferred to a polyvinylidene fluoride membrane. After blocking the non-specific binding sites, membranes were incubated with primary antibodies (Supplementary Table S1) at 4 °C overnight. Subsequently, membranes were washed three times with Tris-buffered saline containing polysorbate 20 (TWEEN 20) and incubated with horseradish peroxide-conjugated secondary antibodies (Cell Signaling Technology, Beverly, MA, USA) at room temperature for 1 h. Chemiluminescence signals were then generated by Western Lightening Plus-ECL (Perkin Elmer, Waltham, MA, USA) and captured by LAS4000 (Fujifilm, Tokyo, Japan). Quantification of protein and phosphorylation levels was performed by ImageJ (1.51 K).