Detection of V5-fLuc after intrastriatal LV injection in mice. As proof of principle experiment, V5 was fused to fLuc in order to facilitate the transgene detection. A) HEK293T cell lysates transduced with LV encoding V5-fLuc-T2A-eGFP were analyzed by SDS-PAGE. The overexpressed fusion protein was detected with both the anti-fLuc antibody (1/3000) and the anti-V5 antibody (1/250000). The co-expressed eGFP was detected by our in-house anti-eGFP antibody (1/10000). B) The same LV as in A) was stereotactically injected in mice striatum. Luciferase activity was confirmed by non-invasive BLI four days and two weeks after injection. A representative image of luciferase activity 4 days post-injection is shown. C) Striatal sections of the same mice as described in B) were immunohistochemically stained with anti-eGFP (1/10000), anti-V5 (1/500) and anti-fLuc antibody (1/500). Scale bar: 0.5 mm.
Lobbestael et al. BMC Biotechnology 2010 10:16 doi:10.1186/1472-6750-10-16