Wave1

Western blots were performed using standard tris-glycine SDS-PAGE protocols. Samples had normalized protein concentration assessed using a Bradford assay (#23246, Thermo). Resolved proteins were electroblotted onto PVDF membranes using semi-dry transfer, blocked for 30 min in TBS supplemented with 0.1% Tween-20 and 5% skim milk, and incubated with primary antibodies (GAP43 #MAB347 Millipore; Actb #A5441 Sigma; Gm130 #610823 BD Biosciences; MAP2 #M1406 Sigma; WAVE1 #ab272916 abcam; WAVE2 #3659T Cell Signaling Technologies; WAVE3 #2806S Cell Signaling Technologies; Cpeb4 #ab224162 abcam; Rbms1 #ab150353 abcam; Tau #AF3494 RR&D) in TBS supplemented with 0.1% Tween-20 and 3% BSA over night at 4°C. The next day, the membrane was washed three times in TBS supplemented with 0.1% Tween-20, incubated for 2 hours at room temperature with secondary antibodies that were isotype-specific, HRP-conjugated, and cross-absorbed (Life Technologies), then washed again three times with TBS supplemented with 0.1% Tween-20. Immunoreactive bands were visualized through detection of chemiluminescence using a CCD camera imager (FluoroChemM, Protein Simple). Quantification of signal intensity in the various bands was performed using the standard Fiji software package.