Journal: Journal of Virology
Article Title: BCA2/Rabring7 Interferes with HIV-1 Proviral Transcription by Enhancing the SUMOylation of IκBα
Figure Lengend Snippet: The BCA2-mediated block on NF-κB delays proviral transcription and replication in CD4 + T cells. Jurkat CD4 + T cells (A) and ConA-activated and IL-2-expanded human PBMCs (B) were transduced with a vector encoding HA-BCA2 or treated with an empty vector (pQCXIP). Forty-eight hours later, the cells were infected with HIV-1 NL4.3. Proviral transcription was assessed at the selected time points by measuring nef RNA levels by RT-qPCR. (Right) The cell lysates derived from these experiments were analyzed by Western blotting for β-actin, BCA2, and p55/p24. (C) HIV-1 NL4.3 replication was also assessed in PBMCs transduced with an empty vector or HA-BCA2 by measuring the amounts of capsid p24 released to the culture supernatant at days 2, 4, 6, 8, and 10 postinfection. (D) The endogenous levels of BCA2 and β-actin were monitored over time in uninfected Jurkat CD4 + T cells and human PBMCs. The data correspond to the mean and standard deviation of three biological replicates, measured in technical replicates. Values that are significantly different are indicated by asterisks (*, P ≤ 0.05; **, P ≤ 0.01; ***, P ≤ 0.001; n.s., not significant).
Article Snippet: Similar replication assays were performed in concanavalin A (ConA)-activated and IL-2-expanded human PBMCs (Zenbio Inc.) transduced with a retroviral empty vector (pQCXIP) or a retroviral vector encoding HA-BCA2.
Techniques: Blocking Assay, Transduction, Plasmid Preparation, Infection, Quantitative RT-PCR, Derivative Assay, Western Blot, Standard Deviation