Figure 2.

Mitochondrial localization of p13^II. (a) Primary rat embryo fibroblasts transfected with the p13^II expression plasmid pSGp13^II using Fugene 6 (Roche) are shown; (b) A human T-cell (Jurkat TetOn cell line; Clontech) transfected with the p13^II expression plasmid pcDNAp13^II using MegaFectin (Q.Biogene) are shown. After formaldehyde-fixation and permeabilization with Nonidet P40, the cells were analyzed by indirect immunofluorescence using a rabbit antibody recognizing p13^II and Alexa 488-conjugated anti-rabbit secondary antibody (Molecular Probes); to visualize mitochondria, the cells were counterstained with mouse anticytochrome c antibody (Pharmingen; a) or goat anti-Hsp60 antibody (Santa Cruz; b) and appropriate Alexa 546 red-conjugated secondary antibodies (Molecular Probes). In addition to verifying colocalization of 13^II with the mitochondrial marker proteins, (a) shows the dramatic changes in mitochondrial morphology induced by the protein