FIG 8.

KPT-335 induces NF-κB nuclear accumulation and varied cytokine expression. RSV A2-infected A549 cells were treated with 1 μM KPT-335, DMSO, or 750 nM LMB from 2 h p.i. Fractions were collected at 24 h p.i. utilizing an NE-PER nuclear and cytoplasmic purification kit, and analyzed by Western blotting as described in the legend of Fig. 6. Blots were probed for p65, and GAPDH and lamin B1 were used as controls. The relative amount of p65 in the nucleus compared to that in the cytoplasm was estimated by densitometric scanning of the protein bands using ImageJ. Data shown are relative to values for DMSO-treated samples, taken as 1. (B) RSV-infected A549 cells were treated with DMSO, KPT-335, or LMB from 2 h p.i.; mock-infected cells served as controls. Supernatants were collected at 24 h p.i. Culture supernatants were clarified by centrifugation before proinflammatory cytokine and chemokine protein expression was examined with a Multi-Analyte ELISArray kit. Fold change was calculated relative to values in mock-infected samples.