Figure 7
From: A role for the histone deacetylase HDAC4 in the life-cycle of HIV-1-based vectors
Effects of overexpression of HDAC4 mutants on stable integration in ATM-deficient cells. (A) Control HeLa cells and HeLa cells overexpressing HDAC4 mutants were infected and treated with the ATM inhibitor. One day post-infection, cells were harvested, DNA extracted and stable integration analyzed by Alu-PCR. C1 - cells transfected with the control EGFP expressing PEGFP-C1 plasmid and infected with the vector, 1061 - cells expressing the HDAC4-1061 mutant and infected with the vector, 1084 - cells expressing the HDAC4-1084 protein and infected with the vector. KU - the ATM inhibitor, KU-55933. +Alu - DNA was analyzed using Alu-PCR, -Alu - a negative control, the Alu primer was left out in the first round of PCR. (B) Comparison of the levels of overexpressed HDAC4 proteins. Western blotting was performed with an anti-GFP antibody (sc-9996, Santa Cruz), since HDAC4-1061 and HDAC4-1084 are fused to the GFP protein [29].