Figure 3
From: Interactions between HIV-1 Vif and human ElonginB-ElonginC are important for CBF-β binding to Vif
EloB significantly stabilizes Vif. 293 T cells (106) were cotransfected with 1 μg of VR-Vif or VR-Vif-SLQ, either without (- Elongin BC) or with co-transfection of 1 μg VR-EloB (+ Elongin B), 1 μg VR-EloC (+ Elongin C), or both 1 μg VR-EloB and 1 μg VR-EloC (+ Elongin BC). Cells were treated with 100 μg/ml CHX 36 h after transfection and harvested at the indicated time points. Western blot analysis of Vif/Vif-SLQ (upper panels) was performed using an anti-Vif antibody. EloB-Myc (EB-Myc) and EloC-HA (EC-HA) were detected with appropriate epitope-tag antibodies (lower panels). (A) Positions of Vif are indicated on the right (upper). (B) Positions of Vif-SLQ are indicated on the right (upper). (C) Individual band intensities of Vif in (A) were quantified using Bandscan software, normalized to the quantity of tubulin bands, expressed as percentages of the value at 0 h (set to 100%) and graphically represented. (D) Quantification of Vif-SLQ bands in (B) was performed as described in (C). All results are representative of three independent experiments. Error bars represent ± SD (n = 3).