Figure 9.

14-3-3 isoforms interact with TOMM34 ex vivo in a phosphorylation-dependent manner. TOMM34^−/− MCF-7 cells transfected with pCMV-N-HA-TOMM34 WT and pCMV-N-HA-TOMM34-S93A/S160A constructs were treated with PMA (0.3 μm), PKA inhibitor H89 (20 μm), and vehicle control (DMSO, -) for 12 h, collected, and lysed. Next, biotinylated 14-3-3 isoforms prebound to streptavidin–agarose beads were incubated with the cell lysates at 4 °C for 1 h. The eluted complexes were analyzed by gel electrophoresis and Western blotting using anti-HA antibody.