Western Blot Analysis of Autophagy Markers

The protocol of the western blot analysis is described in detail elsewhere [9 (link)]. The primary antibodies were as follows: calpain-1 (1 : 1000; Cambridge, MA, USA, ab108400), calpain-2 (1 : 1000; Abcam, ab126600), LAMP1 (1 : 1000; Abcam, ab24170), CTSB (1 : 1000; Millipore, 06480-1), LC3 (1 : 1000; Abcam, ab63817), Beclin-1 (1 : 1000; CST, Technology, Boston, MA, USA, #3495), P62 (1 : 1000; Abcam, ab109012), ATG5 (1 : 500; CST, #12994), TSG101 (1 : 1000; Abcam, ab125011), CD63 (1 : 1000; Abcam, ab271286), V-ATPase (1 : 500; Abcam, ab105937), VAMP7 (1 : 1000; CST, #13920), and β-actin (1 : 500; Proteintech, Chicago, USA, 66009).