Quantification of Newly Synthesized Proteins

Following BONCAT purification, equal volumes of the elution fraction were loaded and separate by SDS-PAGE and analysed via western blotting as previously described (Evans et al., 2019 (link)). The total amount of newly synthesised proteins was quantified using the REVERT total protein stain (LI-COR, 926–11010), with representative proteins for each cluster were detected using the following primary antibodies: α adaptin (ThermoFisher, MA3-061, 1:500), neuron specific enolase (NSE) (Abcam, ab53025, 1:500), AT6V1B2 (Abcam, ab73404, 1:500), PP2A-A (Sigma-Aldrich, 07–250, 1:1000), AUF-1 (Sigma-Aldrich, 07–260, 1:500), and GAPDH (Millipore, MAB374, 1:1000).