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. Author manuscript; available in PMC: 2012 Oct 15.
Published in final edited form as: Gene Ther. 2010 Aug 19;17(12):1476–1483. doi: 10.1038/gt.2010.110

Figure 1. IKKs gene transfer efficiency and decreased nuclear NF-κB in treated muscle.

Figure 1

(A) In vitro gene transfer of IKKα-(dn and wt) and IKKβ-(dn and wt) in AAV constructs. Studies were performed to confirm gene transfer efficiencies of AAV2-IKKα-dn, AAV2-IKKα-wt, AAV2-IKKβ-dn, and AAV2-IKKβ-wt in infected-293 cells 48 hours post-treatment. Western blot analysis by the use of anti-C-terminuses of IKKα and anti-antibodies shows high levels of IKKα-(dn and wt) as well as IKKβ-(dn and wt) expressions (MWs of 85 kD and 87 kD). (B) One month after intramuscular gene transfer with four different AAV2 vectors, the cryosections of GAS muscles treated at 2 months of age and TA muscles treated at 11 months of age were determined the efficiencies of gene transfer by Western blot analysis. The increased levels of IKKα-(dn and wt) and IKKβ-(dn and wt) were found in AAV2-IKKα-dn, AAV2-IKKα-wt, AAV2-IKKβ-dn, and AAV2-IKKβ-wt treated young and old mdxmuscles compared to PBS treated mdx muscle at same age. (C) The electrophoretic mobility shift assay (EMSA) demonstrates the level of nuclear NF-κB in the gastrocnemius (GAS) muscle of both 1 and 2 month-old young mdx mice and the tibialis anterior (TA) muscle of 11 month-old mdx mice treated by AAV2-IKKα-dn, AAV2-IKKα-wt, AAV2-IKKβ-dn, AAV2-IKKβ-wt, and PBS. A total of 45 μg of nuclear extract from each mouse was analyzed at 1 month post -treatment.