Fig 7.

Overexpression of IN binding domain of BRD2 (residues 640 to 801) increases MLV integration. (A and B) HEK293T-based cell lines stably expressing BRD2(640–801) were challenged with MLV-based (A) and HIV-1-based (B) retroviral vectors expressing EGFP (MOI, 1). Cells were harvested at indicated time points for flow cytometry analysis. Overall GFP fluorescence over time is calculated as mean fluorescence intensity × % gated cells. Error bars reflect duplicate measurements in each experiment. (C and D) Integrated proviral vector copies were quantified by qPCR at 16 days postinfection in cells transduced with MLV-based (C) and HIV-based (D) vectors. Values represent the means ± standard errors of the means; n is >3 throughout. Results were analyzed by unpaired t tests. (E) Expression of the BRD2(640–801) fragment and control cell lines used in this experiment was analyzed by Western blotting using anti-FLAG antibody.