Fig 9.

Analysis of dimeric Ad3 fiber knob mutants with increased affinity to DSG2. (A) Competition of Ad3-GFP virus infection on HeLa cells with dimeric affinity-enhanced mutant Y250F and JO-1 (dimeric wt Ad3 fiber knob). The experimental settings are the same as those described for Fig. 3C. (Left) Percentage of GFP-positive cells. (Right) Mean fluorescence intensity. n = 3. The standard deviations were less than 10%. (B) Competition of Ad3-GFP virus infection on HeLa cells by Ad3 knob mutants with enhanced DSG2 binding but without a dimerization domain. A total of 1.5 × 10⁵ HeLa cells were seeded into a 24-well plate. Cells were incubated with the Ad3 knob mutants at increasing concentrations for 1 h at room temperature. One hundred PFU/cell of Ad3GFP virus was then added, and GFP expression was analyzed 18 h later. (C) TEER on colon cancer T84 cells. The experimental settings were the same as those described for Fig. 5A. The TEER at 4 h is shown. n = 3.