Figure 6. Activation of Ras/Raf/Erk pathway is inhibited in IFI6 expressing cells.

Huh7.5 cells treated with antibody to CD81 for the indicated time were lysed in a reducing sample buffer and subjected to Western blot analysis. Lysates were probed for phosphorylated Raf-1 (Ser259). In addition, IFI6 expressing cells were stimulated with EGF and compared to antibody CD81 cross-linked cells to detect the loss of phosphorylation of Raf-1 at Ser259 (panel A). Activation of Raf-1 was analyzed by measuring phosphorylation at Ser338 cross-linking of CD81 or treatment with EGF (panel B). Blots were reprobed with antibody to actin as a loading control. Quantification of protein levels was performed using ImageJ software from three separate experiments and standard deviations are shown as error bars. Note that cropped gel images are used in this figure and the gels were run under the same experimental conditions.