Analysis of ROR1 silencing in MCL cell lines and primary samples. (A) Cell-viability counts of shRNA-expressing MCL cells plus or minus DOX for the indicated time (days). The percentage of viable cells was normalized to (−) DOX for each time point. Data are representative of 3 independent experiments. (B) Immunoblot analysis of shRNA-expressing cell lysates (cytoplasmic and nuclear) plus or minus DOX for 4 and 7 days. (C) Analysis of ROR1 expression by flow cytometry after siRNA nucleofection of primary MCL cells. (D) Immunoblot analysis of NF-κB p65 and Wnt5a levels in primary cell lysates. Quantifications shown for NF-κB p65 and Wnt5a protein levels were normalized to β-tubulin levels and compared with siCtr for each sample. (E) Luciferase activity (fold induction) of Mino cells cotransfected with ROR1-HA and NF-κB p65 reporter plasmids for 24 hours. Anti-HA immunoblotting shows ROR1 protein levels. Error bars represent mean plus or minus SD of triplicates. Statistical significances compared with sample without ROR1-HA transfection were calculated by a 2-tailed paired Student t test; *P < .05. Ctr, control; HA, hemagglutinin; PE, phycoerythrin.