Detection of autophagy-related proteins and lysosomes. Autophagy-related proteins, Beclin-1 and microtubule associated protein 1 light chain 3 (LC3) were assessed by the immunofluorescence method. (a) LC3 was stained by Alexafluor 594-labeled second antibody. (b) Beclin-1 was stained with Alexafluor 488-labeled second antibody. (c) The activity of lysosomes was assessed using a Lyso-Tracker kit. The activated lysosomes were shown in the cytoplasm. The annulus fibrosus (AF) cells cultured in 10% fetal bovine serum (FBS) were used as controls.