Atg4b 15 131 1 ap

The FDA-approved Drug Screening Library (L1000) was from TargetMol (Boston, MA). Candidate compound 10 (M832611), 424 (B802495), 503 (H811537), 523 (L830883), 602 (O815322), 836 (R830529), 1574 (S832949) were purchased from Macklin (Shanghai, China) and compound 669/Ebselen (E3520) and 1358 (232,120) were from Sigma (St. Louis, MO). S130 [10 (link)] and FMK-9a [23 (link)] were preserved in our lab. Z-VAD-FMK (S7023) and Staurosporine (STS, S1421) were from Selleckchem (Houston, TX). Chloroquine (CQ, A506569) was from Sangon Biotech (Shanghai, China). The sources of different atomic analogues of Ebselen were as follows: Se atom analogues of Se-1 ~ 4 were a gift from professor Kewu Yang (Northwest University), and other O, SO, S atom analogues were from BidePharm (Shanghai, China).
The antibody ATG4B (15,131-1-AP) and GAPDH (60,004-1-Ig) was from Proteintech (Wuhan, China), LC3B (L7543) was purchased from Sigma (St. Louis, MO), antibody anti-LC3B (PM036) for immunofluorescence was from MBL (Beijing, China), Secondary antibodies conjugated with peroxidase and Alex Fluor-488 were from ThermoFisher Scientific (Waltham, MA).

Antibodies used in this study were as follow: ATG4B (15131-1-AP) was from Proteintech (Wuhan, China), ATG5 (8540) and FIP200 (12436) were from Cell Signaling Technology (Danvers, MA, USA), GAPDH (sc-365062) was from Santa Cruz Biotechnology (Dallas, TX, USA), LC3B (L7543) and TUBA (T6074) were from Sigma (St. Louis, MO, USA), LAMP1 (H4A3) was from Developmental Studies Hybridoma Bank (Iowa City, IA, USA), SQSTM1/p62 (PM045) was from MBL International (Woburn, MA, USA). Secondary antibodies (35503, 35511, 35553, and 35561) were purchased from Thermo Fisher Scientific (Rockford, IL, USA).
Bafilomycin A1 (Baf, B-1080) was purchased from LC Laboratories (Woburn, MA, USA), as well as rapamycin (Rap, ASW-125). Chloroquine (CQ, A506569) was purchased from Sangon Biotech (Shanghai, China). Z-VAD-FMK (S7023) and staurosporine (SP, S1421) was from Selleckchem (Houston, TX, USA). Dequenched-BSA (DQ-BSA) Red (D12051), LysoTracker Red (LTR, L12492) and Lipofectamine 2000 (11668-019) were purchased from Life Technologies (Bothell, WA, USA). Substrate Ac-DEVD-AFC (CASP-048) for caspase3 activity was from Chinese Peptide Company (Hangzhou, China). Purified ATG4B, substrate ECFP-GATE-16-YFP (FRET-GATE16) for ATG4B were described before [45 (link)].