Figure 2.

Cells were incubated with PorB as described in the text. (A) Cytosol and whole membranes from HeLa (lanes 1 and 2) and splenic murine B cells (lanes 3 and 4) were analyzed by Western blot with anti-PorB rabbit polyclonal serum. (B) Mitochondria (m) and cytosolic fractions (c) of CH-12 RMC (lanes 2–5), Jurkat (lanes 6–9), and splenic murine B cells (lanes 10–13) were analyzed with anti-PorB mAb. Purified PorB was used as molecular weight control (lane 1). (C) Mitochondria were isolated from CH-12 RMC and Jurkat cells, and the association of PorB with the purified mitochondria was detected by flow cytometry with anti-PorB rabbit polyclonal serum and FITC-labeled anti-rabbit second antibody. Preimmune rabbit serum was used as negative control for nonspecific binding, which was negligible.