Figure 7.

Transduction ability of alanine mutant capsids. HEK293T cells were transduced with viruses packaging scGFP transgene at a multiplicity of infection of 1000, and transgene expression was quantified at 48 h post-transduction using flow cytometry. (A) Transduction index (TI) of viruses. TI is the product of percentage of GFP+ cells and the geometric mean fluorescence intensity; metrics were obtained from flow cytometry. N = 3 independent experiments done in triplicate. Error bars are mean ± SE. One-way ANOVA was performed with Dunnett’s post hoc multiple comparison test to compare mutants with WT. TI of alanine mutants is plotted against (B) number of residue-residue contacts in the native capsid structure, (C) frequency of alanine in the parvovirus sequence family at the site of the mutated residue, (D) Δ-frustration index, and (E) ΔH_DCA(Ala) score. N = 8 mutants for structure-based analyses. N = 6 mutants for sequence-based analyses because R471A and S721A were excluded from these analyses because these residues do not exist in a majority of the sequence family (these residues are considered as insertions in the hidden Markov model of the family). Regression lines with 95% confidence interval are plotted for statistically significant correlations. R²-values and significance levels are reported. ^∗p < 0.05, ^∗∗p < 0.01, ^∗∗∗p < 0.001.