Scrambled control shrna vector

MM cell lines and primary cells were infected with lentiviral vectors expressing a MUC1 shRNA, NF-κB p65 shRNA or scrambled control shRNA vector (Sigma) [8 (link), 46 (link)]. Cells were selected and maintained in puromycin. For DOX-inducible MUC1 and MYC silencing, MUC1shRNA (MISSION shRNA; Sigma, TRCN0000122938), MYCshRNA (MISSION shRNA; Sigma, TRCN0000039642) or a control scrambled CshRNA (Sigma) was inserted into the pLKO-Tet-puro vector (Addgene, Cambridge, MA, USA, Plasmid #21915). Viral vectors were produced in HEK293T cells as described [51 (link), 52 (link)]. Cells expressing Tet-shRNA vectors were selected in puromycin. MUC1 was also silenced with CRISPR/Cas9 gene editing as described [8 (link)].

Human THP-1 (ATCC) and MOLM-14 (ATCC) cells were cultured in RPMI1640 medium supplemented with 10% heat-inactivated fetal bovine serum, 100 units/ml penicillin, 100 ug/ml streptomycin and 2 mM L-glutamine. Authenticity of the cells was confirmed by short tandem repeat (STR) analysis [44 (link)]. Cells were infected with lentiviral vectors expressing a MUC1 shRNA, NF-κB p65 shRNA or scrambled control shRNA vector (Sigma). Cells were selected and maintained in puromycin [40 (link)]. Cells were treated with the NF-κB pathway inhibitor BAY-11-7085 (Santa Cruz Biotechnology) or DMSO as a vehicle control.