BLyS ligand stimulation increases BAFF-R nuclear localization. (A) Normal isolated peripheral blood B cells were stimulated with anti-IgM, BLyS, or both and then probed with BAFF-R (Cy3) and TOPRO-3. Samples were then analyzed by confocal microscopy. (B) WB of nuclear extracts from normal peripheral blood B cells, treated with anti-IgM, human recombinant BLyS, or both, probed with a BAFF-R antibody. Lamin B was used as a nuclear protein loading control. (C) WB of cytoplasm extract from NHL-B cells (MS) transfected with BLyS siRNA (siBLyS) or scrambled siRNA, and probed with BLyS antibody (left panel). Actin was used as loading control. WB of nuclear extracts from NHL-B cells (MS) transfected with BLyS siRNA or scrambled siRNA and probed with BAFF-R antibody. Lamin B was used as a nuclear protein loading control (right panel).