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. 2007 Nov 7;82(2):630–637. doi: 10.1128/JVI.01896-07

FIG. 3.

FIG. 3.

Validation of the affibody-modified fiber replacement proteins. (A) Western blot analysis of the transiently expressed proteins in cell lysates containing the wt Ad5 fiber (lanes 1 and 2), 11F.dZ (lanes 3 and 4), 11F.Her2:7 (lanes 5 and 6), or F11.Her2:4 (lanes 7 and 8) in seminative form (odd-numbered lanes) or denatured form (even-numbered lanes). Each line of the gel was loaded with the aliquot of cell lysate containing 8 μg of total soluble protein. S, protein molecular mass standards. (B) Fluorescence-activated cell sorting assay of the transiently expressed fibers: wt Ad5 fiber (green), 11F.dZ (orange), 11F.Her2:4 (blue), and 11F.Her2:7 (red). The black curve represents the background signal obtained with the lysate of mock-transfected cells. (C) Dose-dependent inhibition of cell attachment of the 11F.Her2:7 protein detected by flow cytometry. Mean fluorescence intensity of the 293/Her2 cells probed with either the 11F.Her2:7-containing lysate (black bars) or the wt fiber-containing lysate (white bars). Shown below the graph are the concentrations of the free affibody competitor (in mg/ml) used to inhibit Her2 binding. Error bars indicate the standard deviations obtained in triplicate samples.