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. 2012 Apr;86(8):4527–4537. doi: 10.1128/JVI.07071-11

Fig 1.

Fig 1

Adenovirus activation of RAW 264.7 cells. (A) RAW 264.7 cells infected with the indicated amount of virus (particles/cell) were harvested at 2, 4, and 6 h postinfection as described in Materials and Methods and analyzed by Western blotting (probed with pser388IRF3-, IRF3-, or actin-specific antibody). (B) The IRF3 activation assay was performed as described for panel A, comparing cells exposed to 20,000 p/cell of Ad5CiG or bacterial endotoxin LPS (100 ng/ml). A Western blot was probed with phosphospecific antibodies pser388IRF3, ptyr701STAT1, and ptyr689STAT2 as well as IRF3- and actin-specific antibodies as total protein controls. (C) RAW 264.7 cells were transfected with Ad5CiG DNA or poly(I·C), and cell lysates were harvested and analyzed by Western blotting as previously described. (D) IRF3 activation assay comparing cells exposed to 20,000 p/cell of Ad5CiG or the endosomal escape mutant Ad2ts1. Samples were harvested at 4 and 6 h postinfection and analyzed by Western blotting as previously described. All experiments have been done a minimum of three times.