Fig. 5

Characterization of the effect of ECM depletion on RGD4C.AAVP-guided gene transfer in 9L cells. a Luciferase expression in 9L cells by Steady-Glo® assay after treatment with increasing concentrations of collagenase or hyaluronidase, at day 3 post-transduction with RGD4C.AAVP/Luc vector carrying the Luc reporter gene. b Time course expression of luciferase over 5 days post transduction with RGD4C.AAVP vector alone, or RGD4C.AAVP in conjunction with collagenase (0.2 mg/ml) or hyaluronidase (0.4 mg/ml) or with combination of both enzymes. Similar enzymatic treatments were included with the control non-targeted NT.AAVP vector. c GFP expression in 9L cells transduced with RGD4C.AAVP-GFP alone (control) or following various ECM depletion strategies: collagenase, hyaluronidase or combination of both enzymes. Images were visualized by fluorescence confocal microscopy 3 days post vector transduction One way ANOVA was used, together with Tukey’s post-test to generate the data, *, p < 0.05, **, p < 0.01, ***, p < 0.001. Luciferase expression results are shown as relative luminescence units/1 μg protein. Samples were obtained through triplicate wells and each experiment was performed at least twice with consistent results. Data are presented as mean ± s.e.m. Experiments were repeated twice and individual conditions were repeated in triplicates