Puromycin incorporation assay (SUnSET assay)

The SUnSET assay was performed according to previously published protocol (Schmidt et al, 2009), with modifications. Briefly, neurospheres were treated with 10 µg/mL puromycin for 1 h. Puromycin-incorporated proteins were detected by western blotting using an anti-puromycin antibody. Ponceau S staining was used as a loading control.

Reference

Schmidt EK, Clavarino G, Ceppi M, Pierre P (2009) SUnSET, a nonradioactive method to monitor protein synthesis. Nat Methods 6:275–277