Mg132 bml pi102

Urea, DTT, and buffer reagents were purchased from Sigma-Aldrich (St. Louis, MO, USA). Purified ubiquitin (BML-UW8795), ubiquitin chains (BML-UW0825), and MG-132 (BML-PI102) were obtained from Enzo Life Sciences (NY, USA).

Slug (9585S), CUL1 (4995S), NEDD8 (2745S), p-AKT (ser473) (9271S), phospho-mTOR (ser2448) (2971S), E-Cadherin (3195S), Phospho-STAT3 (9145S), Phospho-Smad2/3 (8828S), Phospho-ERK (9101S), and antibodies of EMT markers (ZO-1, Vimentin, ZEB1, N-cadherin, Claudin-1, β-Catenin, Snail) (Epithelial–Mesenchymal Transition (EMT) Antibody Sampler Kit 9782T) were purchased form Cell Signaling Technology (Danvers, MA); β-tubulin (sc-166729), p53 (sc-126), p21 (sc-6246), MDM2 (sc-965), and Ubiquitin (sc-8017) antibodies from Santa Cruz Biotechnology (Dallas, TX). MLN4924 was synthesized, as previously described [26 (link)]. MG132 (BML-PI102) (Enzo Life Sciences, Plymouth Meeting, PA, USA), Cycloheximide (C1299) (Sigma Aldrich, St. Louis, MO, USA), LY294002 (L9908), MK-2206 (s1078), and Rapamycin (s1039) (Selleck Chemicals, Houston, TX, USA) were purchased from the indicated company.

All reagents unless otherwise noted were from Sigma. FluoZin™-3, AM (FZ3; zinc probe) from ThermoFisher (F24195) was reconstituted with DMSO to a stock solution of 500 μM. Lectin PNA (A. hypogea/peanut agglutinin) conjugated to Alexa Fluor™ 647 (PNA-AF647) was from Invitrogen™ (L32460). Fluo-4 NW (calcium probe) from ThermoFisher (F36206) was reconstituted using kit provided assay buffer. Hoechst 33342 (H33342) from Calbiochem (382065) was reconstituted with H₂O to a stock solution of 18 mM. PI from Acros Organics (AC440300010) was reconstituted with H₂O to a stock solution of 1 mg mL⁻¹. Proteasomal inhibitors were from Enzo Life Sciences: MG132 (BML-PI102) was reconstituted with DMSO to a stock solution of 20 mM; epoxomicin (Epox, BML-PI127) was reconstituted to a stock solution of 20 mM (using MG132 stock); and clasto-lactacystin β-Lactone (CLBL, BML-PI108) was reconstituted with DMSO to a stock solution of 5 mM. Zn-chelator TPEN from Tocris (16858-02-9) was resuspended with 1:100 EtOH:H₂O to a stock solution of 1 mM. Bovine serum albumin was from Sigma (A4503). Anti-phosphotyrosine antibody, clone 4G10^® was from EMD Millipore (05-321).