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. 2010 Nov 5;286(1):777–786. doi: 10.1074/jbc.M110.193441

FIGURE 4.

FIGURE 4.

14-3-3β binding prevents apoptotic SRPK2 cleavage. A, SRPK2 mutant apoptotic cleavage assay. HEK293 cells were transfected with Myc-tagged SRPK2WT, T492A, or T492D for 24 h, and then the cell lysates were incubated with active caspase-3 at 37 °C for 2 h. B, 14-3-3β wild-type but not K50E mutant protects SRPK2 from apoptotic cleavage. HEK293 cells were transfected with Myc-tagged SRPK2WT, T492A, or T492D and 14-3-3WT or 14-3-3K50E for 24 h, and then the cell lysates were incubated with active caspase-3 at 37 °C for 2 h. C, 14-3-3β protects SRPK2 from VP16-triggered apoptotic cleavage. HEK293 cells were transfected with Myc-tagged SRPK2WT, T492A, or T492D and 14-3-3WT or 14-3-3β-K50E for 24 h, and then treated with DMSO or VP16 for another 16 h. The cell lysates were analyzed by immunoblotting. D, D139A mutation affects SRPK2 binding to 14-3-3β. HEK293 cells were co-transfected with Myc-tagged SRPK2WT, T492A, or T492D and GFP-14-3-3β-WT for 48 h, and then the cell lysates were subjected to immunoprecipitation with anti-14-3-3β antibody. The coprecipitated proteins were analyzed by immunoblotting with Myc antibody.