Skip to main content
. 2003 Dec 5;101(1):153–158. doi: 10.1073/pnas.2533920100

Fig. 6.

Fig. 6.

Multisite phosphorylation of Bcl2 induced by either paclitaxel or substitution of glutamate can enhance Bcl2's stability and resistance to proteolysis. (A) Cells expressing WT or various A- or E-Bcl2 mutants were metabolically labeled with [35S]methionine. The t1/2 of WT and mutant Bcl2 was determined by classic pulse–chase analysis. (B and C) Mobility-shifted Bcl2 was immunoprecipitated from lysates of cells expressing WT Bcl2 after treatment with paclitaxel to induce multisite Bcl2 phosphorylation. A- and E-containing Bcl2 mutants were immunoprecipitated from lysates of untreated cells expressing A- or E-mutants. These Bcl2 immune complexes were then incubated with an apoptotic extract for 2 h, and Bcl2 was analyzed by Western blotting by using Bcl2 antibody. (D) NSF.N1/H7 cells expressing equivalent amounts of WT or A- or E-Bcl2 mutants were treated with 1 μM staurosporine for 24 and 48 h. Cell viability was assessed as described for Fig. 1D. Data represent the mean ± SD of three independent determinations.