we developed a novel method combining axon-pSILAC17 with axonal ribosome purification. In this method, we first labeled newly synthesized proteins with heavy amino acids (AAs) in somaless axons cultured in a Boyden chamber. Approximately 2000 eyes were cultured for each experiment to obtain sufficient axonal material. Then, axonal ribosomes were purified using a sucrose cushion and ultracentrifugation, followed by identification of newly synthesized proteins with mass spectrometry.