Figure 5. SLC35A1 KO leads to a larger decrease in transduction efficiency than that of vector binding and entry of rAAV5 or rAAV2.5T in polarized HAE-ALI cultures.
(A-C) rAAV5 vector binding, internalization, transduction in HAE-ALISLC35A1-KO cultures. HAE-ALI Scramble, SLC35A1 KO or NA-treated scramble cultures were apically transduced with rAAV5 at an MOI of 20,000 DRP/cell. At 2 hpt, vector binding and internalization assays were carried out, and at 5 dpt, the luciferase activities were measured. Relative percentage of binding (A), internalization (B) or transduction efficiency (C) of the transduced KO and NA-treated cultures to the Scramble cell group are shown. (D-F) rAAV2.5T vector transduction, binding, and internalization in HAE-ALISLC35A1-KO cultures. The ALI cultures as indicated were transduced with rAAV2.5T at an MOI of 20,000 DRP/cell. Relative percentages of vector binding (D), internalization (E), and transduction efficiency (F) of the transduced KO and NA-treated cell cultures to the Scramble cell group are shown. All repeated data are shown with a mean and SD of at least three replicates. P values as indicated were determined by using one-way ANOVA for the comparison of the vector value in the KO or NA-treated cell group with the Scramble cell group.