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Figure 5 | Retrovirology

Figure 5

From: Caspase-3-mediated cleavage of p65/RelA results in a carboxy-terminal fragment that inhibits IκBα and enhances HIV-1 replication in human T lymphocytes

Figure 5

Dimerization of ΔNH 2 p65 in Jurkat cells. (a) Schematic representation of ΔNH2p65-tag mutant, which carries the ATG codon at Asp97. This mutant lacks part of DNA contact domains but not the dimerization domain. (b, c) Ten micrograms of cytosolic extracts from Jurkat cells transiently transfected with either pCMV-p65wt-tag, pCMV-p65 D94E;D97E-tag or pCMV-ΔNH2p65-tag expression vectors were analyzed by immunoblotting using an antibody against the carboxy-terminus of p65/RelA (b) and anti-FLAG tag M2 mAb (c). Untagged plasmid pCMV-p65wt was used as a control of the anti-FLAG tag M2 mAb specificity. (d) Two hundred micrograms of protein extracts from Jurkat cells transiently transfected with pCMV-p65wt-tag, pCMV-p65 D94E;D97E-tag and pCMV-ΔNH2p65-tag expression vectors were subjected to immunoprecipitation using the anti-FLAG tag M2 mAb. Analysis was carried out by immunoblotting using antibodies against the carboxy-terminus of p65/RelA, NF-κB1/p50 and IκBα. Images correspond to the same western blot gel that was first blotted simultaneously with antibodies against p65/Rel and IκBα and then it was deshybridized and reprobed with anti-NF-κB1/p50.

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Retrovirology

ISSN: 1742-4690

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