Ampho envelope vector

Wild-type or gene-deficient C57BL/6 mice were housed at NII and used in accordance with the IAEC guidelines. MEFs, immortalized using 3T3 protocol, were generated from E13.5 embryos, as described.^{32 (link)} HMCLs were a kind gift from Dr Michael Kuehl, NCI. Indicated transgenes were either constitutively expressed from MoMLV LTR promoter from pBabe.puro or inducibly expressed from an engineered promoter containing five tandem kappaB sites from HRS.puro retroviral construct. Retrovirus particle were generated from 293 T cells co-transfected with pCL-Eco.^{32 (link)} For gene transduction into human-derived myeloma cells, pseudotyped retrovirus particles were produced using Retro-X Universal Packaging System (Clonetech, Mountain View, CA, USA) from GP2-293 cells co-transfected with pBabe.puro construct and Ampho envelope vector (Clonetech). Following shRNA-expressing lentiviral particles (Sigma Aldrich, St Louis, MO, USA) were used for knockdown in HMCLs: Rela#1-TRCN0000353629, Rela#2-TRCN000029875, Relb#1-TRCN0000280360, Relb#2-TRCN0000280361, Nfkb2#1-TRCN0000356047 and Nfkb2#2-TRCN0000356005, and control-SHC202V. For knockdown studies in fibroblasts, lentiviral constructs expressing control shRNA (#RHS4346) or RelB-shRNAs (RMM#4431-101264262 and RMM#4431-200408572) purchased from GE-Dharmacon (Lafayette, CO, USA) were used for generating lentiviral particles.^{49 (link)}