Myc taq

Dulbecco's modified Eagle's medium (DMEM, LM001-05), fetal bovine serum (FBS, S001-07) and Roswell Park Memorial Institute (RPMI, LM011-01) 1640 medium were purchased from Welgene (GD, Korea). 3-methyladenine (3-MA, 5142-23-4), Z-Leu-Leu-Leu-al (MG132, C221) and cycloheximide (CHX, 01810) were purchased from Sigma-Aldrich (St Louis, MO, USA). X-tremeGENE HP DNA transfection reagents (06366236001) were purchased from Roche (Mannheim, Germany). DC Protein Assay Kit II (500-0113, 0114 and 0115) was purchased from Bio-Rad (Hercules, CA, USA). ECL western blotting detection reagents (RPN2209) were purchased from GE Healthcare (Buckinghamshire, UK). Antibodies for CNOT2 (34214, 1:2000 dilution), p62/SQSTM1 (5114, 1:2000 dilution), Myc-taq (2276, 1:2000 dilution), LC3B (3868, 1:1000 dilution) and ATG5 (8540, 1:2000 dilution) were purchased from Cell Signaling Technology (Beverly, MA, USA). Antibodies for OctA-taq (Flag, SC-807, 1:2000 dilution), HA-taq (SC-7392, 1:2000 dilution), immunoglobulin G (IgG, SC-69786), actin (SC-47778, 1:5000 dilution), 4′, 6′-diamidino-2′-phenylindole dilactate (DAPI, 28718-90-3) and Protein A/G plus-agarose (SC-2003) were purchased from Santa Cruz Biotechnology (Dallas, Texas, USA). Other chemical reagents were obtained from Sigma-Aldrich.

Cycloheximide, 5-aza-2′-deoxycytidine (5-Aza), trichostatin A (TSA), and rapamycin were purchased from Sigma-Aldrich (St. Louis, MO, USA). The indicated primary antibodies against the following proteins were used in this study: STAT1/2/3/4/5/6 (#9172/4594/12640/5097/9363/9362), DNMT1/3a/3b (#5119/3598/67259), cleaved-caspase 3/7 (#9664/9491), cleaved PARP (#5625), S6K (#2708), phospho-S6K (T389) (#9205), S6 (#2217), phospho-S6 (Ser235/236) (#2211), mTOR (#2972), 4E-BP1 (#9644), phospho-4E-BP1 (T70) (#9455), eIF4E (#2067), phospho-eIF4E (S209) (#9741), eIF4G (#2469), Rheb (#13879), TSC2 (#4308), p-TSC2 (T1462) (#3617), p-TSC2 (S1387) (#5584), ICAM1 (#4915), JAK2 (#3230), NFATC2 (#4389) and Myc taq (#2278) (Cell Signaling Technology, Danvers, MA, USA); anti-HIF-1α (610958) (BD Biosciences, San Jose, CA, USA); and anti-actin (sc-1616) and GAPDH (sc-48,167) (Santa Cruz Biotechnology, Santa Cruz, CA, USA). Secondary antibodies used were anti-goat IgG HRP (81–1620; Invitrogen, Carlsbad, CA, USA), anti-mouse IgG HRP (G-21040; Invitrogen), and anti-rabbit IgG HRP (111–035-003; Jackson Laboratories, Bar Harbor, ME, USA).