Figure 5. Depletion of PAK1 enhanced Survivin-ubiquitination in pancreatic β-cells. (A) MIN6 cells were transiently transfected with siPAK1 or siCon Oligos. Thirty-six hours after transfection, cells were cultured with proteasome inhibitor MG132 (10 µM) for 6 h. Whole cell detergent lysates were prepared and subjected to 12% SDS-PAGE for immunoblotting with antibodies indicated. Data represent the one of the two independent experiments yielding similar results. (B) MIN6 cells were treated as in A. One mg cleared detergent lysates were combined with 1 µg of rabbit anti-Ubiquitin antibody as described in methods. The immunoprecipitates were subjected to immunoblotting with anti-survivin antibody. Data represent the one of the three independent experiments yielding similar results.