Figure 6.

Kinase inhibitors effects on AAV2 entry, capsid phosphorylation, and ataxia-telangiectasia mutated (ATM) activation. (A) Representative WB analysis (n = 2) of immunoprecipitates (IP) from lysates of HEK293 cells pretreated with the various drugs and infected with dual AAV2 vectors expressing eGFP. Immunoprecipitates were blotted with anti-β1 integrin (α-β1-Int), anti-phospho-tyrosine (α-P-Tyr), or anti-phospho-serine (α-P-Ser) antibodies indicated on the left. Parallel samples of the same immunoprecipitates were blotted using the anti-VP (α-VP) antibodies. Molecular weight markers are shown on the left. IP, immunoprecipitation; arrows point at β1 integrin and AAV VP3. (B) Phospho (Ser 1981) ATM measured by enzyme-linked immunosorbent assay in lysates of HEK293 cells infected for 2 h with dual AAV2 and incubated for 1 h with the kinase inhibitors. Values (n = 3) are expressed as the mean percentage of the values measured in cells not infected and incubated with DMSO ± SE. Drug concentrations: F13 5 μM, H17 10 μM, and K17 1 μM. *p < 0.05.