FIG 1.

Generation and characterization of NF-κB-defective MLE-15 cells. (A) Cells were transfected with the vector px459 or px459-mp65. Transfected cells were selected by puromycin treatment, and surviving clones were grown to colonies. A fraction of the cells was lysed, and equal amounts of protein were tested by immunoblotting for expression levels of p65, Cas9, and tubulin with specific antibodies. (B) The experiment was done as in panel A, with the difference that cells were transfected with px459-mNEMO and extracts from cell clones were tested for the expression of NEMO. (C) The indicated cells were infected with SC35 or SC35M (MOI of 1), and IL-6 gene expression was quantified by qPCR 24 h p.i. Error bars display standard errors of the means (SEM) derived from two independent experiments performed in triplicate. Student's t test was used for statistical analysis. **, P < 0.01. All other differences had P values of <0.001. ns, not significant.