Fig. 2: BBRF2 interacts with BSRF1.
a Co-immunoprecipitation of BBRF2-Myc and BSRF1-Flag in HEK293T cells. The Flag vector was used as a negative control. IB immunoblotting; Input, whole cell lysis. Source data are provided as a Source Data file. b Confocal immunofluorescence imaging of BBRF2 (green) and BSRF1 (red) in HeLa cells. Cell nuclei were counterstained with DAPI (blue). Scale bars, 5 μm. Each experiment was repeated three times independently with similar results. c The binding affinity of BBRF2Δ with BSRF1Δ measured by Biacore. His-tagged BBRF2Δ (2.5 μg ml−1) was immobilized on the NTA chip and the association signals of BSRF1Δ at different concentrations (12.5−500 nM) monitored. Source data are provided as a Source Data file. d SEC-RALS analysis showing BBRF2Δ and BSRF1Δ form heterodimers in solution, analyzed on a Superdex 75 SEC column. Calculated molecular masses at the absorption peak of 280 nm are plotted in black. mAU, milli-absorption units. e The constructs of full-length BSRF1, BSRF120−218, and BSRF1Δ. f, g SEC-RALS analysis showing that BSRF120−218 is dimeric in solution (Superdex 75), and BBRF2Δ and BSRF120−218 form heterotetramer in solution (Superdex 200). h SEC analysis of the BBRF2Δ-BSRF120−218 complex (blue), BSRF120−218 dimer (pink) and BBRF2Δ (green), with BSA (grey dashed line) as a reference (Superdex 200).
