Fetal calf serum (fcs)

Primary DEF cells were prepared from 12-day-old embryonated Pekin duck eggs by a standard method [32 (link)] and maintained in growth medium consisting of DMEM (Macgene, Beijing, China) supplemented with 10% FCS (Corning, Corning, NY, USA), 100 U/mL of penicillin, and 0.1 mg/mL of streptomycin (10% FCS DMEM). The cells were grown in T25 flasks, seeded at 1 × 10⁶ cells per flask, or 24-well plates, seeded at 2 × 10⁵ cells per well, and incubated at 37 °C in a 5% CO₂ atmosphere.
Two DHAV-1 strains were used: C-QYD, originally isolated in 10-day-old embryonated duck eggs from an outbreak of DVH and passaged once [33 (link)]; and C80, the 80th passage of the DHAV-1 C strain in specific pathogen-free (SPF) embryonated chicken eggs [12 (link)]. The viruses, present as embryo homogenates, were stored at −80 °C. Using embryonated duck and chicken eggs, we determined the titers of C-QYD and C80 to be 10^5.50 and 10^6.84 50% embryo lethal dose (ELD₅₀) per 0.1 mL, respectively.
Antiserum against DHAV-1 C-QYD was prepared in our laboratory previously and stored at −20 °C [34 ].

Sulforaphane was purchased from LKT Laboratories (St. Paul, MO). Bovine serum albumin (BSA), menadione, dicoumarol and NADPH were purchased from Sigma-Aldrich Corporation (St. Louis, MO). Primary antibodies against NQO1, Keap1, Nrf2, NFκB p50, NFκB p65, IκB, caspase 2, caspase 3, bcl-2, bax, actin, histone H1 and secondary antibodies were purchased from Santa Cruz Biotechnology, Inc (Santa Cruz, CA). Anti-poly (ADP-ribose) polymerase (PARP) antibody was purchased from Biomol International, L.P. (Plymouth Meeting, PA). Fetal calf serum, RPMI-1640, penicillin and streptomycin were purchased from Cellgro, Inc (Herndon, VA). All other chemicals and solvents used were of analytical grade.

The Nalm6 [25 (link)] and 697 [11 (link)] have been previously described. CCRF-CEM (CEM), MOLT-4 and U-937 cells were obtained from the American Type Culture Collection (ATCC, Manassas, VA). Cell lines were cultured in RPMI 1640 medium (Cellgro, USA) supplemented with 10% fetal bovine serum (Hyclone, USA). HEK 293T cells were cultured in DMEM (Cellgro) supplemented with 10% fetal calf serum and 1% L-glutamine (Cellgro). Cells were incubated at 37°C in a humidified atmosphere of 5% CO₂. Primary human B-ALL and T-ALL cells were cultured in RPMI 1640 medium (Cellgro) supplemented with 10% fetal bovine serum (Hyclone). 4, 5, 6, 7-Tetrabromobenzotriazole (TBB) was purchased from Sigma (St. Louis). Cells were cultured with or without TBB and collected for total RNA isolation. Human HA-tagged IKZF1 retroviral construct and retroviral production was described previously [14 (link), 16 (link), 26 (link), 27 (link)].

Nalm6 and MOLT4 cells were obtained from the American Type Culture Collection (ATCC, Manassas, VA) and cultured in RPMI 1640 medium (Cellgro) supplemented with 10% fetal bovine serum (Hyclone). HEK 293T cells were cultured in DMEM (Cellgro) supplemented with 10% fetal calf serum and 1% L-glutamine (Cellgro). Cells were incubated at 37°C in a humidified atmosphere of 5% CO₂. Primary human B-ALL and T-ALL cells were cultured in RPMI 1640 medium (Cellgro) supplemented with 10% fetal bovine serum (Hyclone). Cells were cultured with or without TBB and collected for total RNA isolation. Human HA-tagged Ikaros (IKZF1) retroviral construct and retroviral production was described previously [11 (link), 12 (link), 34 (link)].

The Nalm6 cell line has been previously described [33 (link), 34 (link)]. Cells were cultured in RPMI-1640 medium (Cellgro, Tewksbury, MA, USA) supplemented with 10% fetal bovine serum (Hyclone, Logon, Utah, USA). HEK 293T cells were cultured in DMEM (Cellgro) supplemented with 10% fetal calf serum and 1% L-glutamine (Cellgro). Cells were incubated at 37°C in a humidified incubator with 5% CO₂. Primary human B-ALL cells were cultured in RPMI-1640 medium (Cellgro) supplemented with 10% fetal bovine serum (GE-Hyclone, Logon, Utah, USA). CX4945 was purchased from Sigma (St. Louis, MO, USA). Cells were cultured with or without CX-4945 and collected for total RNA isolation. Human IKZF1 retroviral construct and retroviral production was described [33 (link)–35 (link)].

The Nalm6 cell line is previously described46 (link). The CCRF-CEM (CEM), MOLT-4 and U-937 cell lines were obtained from the American Type Culture Collection (ATCC, Manassas, VA). Cells were cultured in RPMI-1640 medium (Cellgro, Tewksbury, MA, USA) supplemented with 10% fetal bovine serum (Hyclone, Logon, Utah, USA). HEK 293T cells were cultured in DMEM (Cellgro) supplemented with 10% fetal calf serum and 1% L-glutamine (Cellgro). Cells were incubated at 37 °C in a humidified atmosphere with 5% CO₂. Primary human B- and T-cell ALL cells were cultured in RPMI–1640 medium (Cellgro) supplemented with 10% fetal bovine serum (GE-Hyclone, Logon, Utah, USA). 4,5,6,7-Tetrabromobenzotriazole (TBB) and CX-4945 were purchased from Sigma (St. Louis, MO, USA). Cells were cultured with or without TBB and collected for total RNA isolation. Human IKZF1 retroviral construct and retroviral production was described15 (link)17 (link)47 (link)48 (link).