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. 2005 Dec;25(23):10543–10555. doi: 10.1128/MCB.25.23.10543-10555.2005

FIG. 10.

FIG. 10.

Hyperosmotic stress regulates the interaction of PKCζ and caspase 9 (Casp-9) in cells. HEK293 cells were transfected with FLAG-PKCζ and His-caspase 9 (C287A) or caspase 3 (C163A). Cells were serum starved for 24 h prior to treatment with NaCl. FLAG-PKCζ immunoprecipitates (IP) and total lysates were analyzed by Western blotting with antibodies against caspase 9, caspase 3, and PKCζ/λ. (A) Coprecipitation of PKCζ with caspase 9 but not caspase 3 and disruption of the interaction between PKCζ and caspase 9 by treatment with 0.7 M NaCl for 40 min. Twenty micrograms of total lysate from samples 4 and 6 was also loaded to estimate the proportions of caspase 9 and caspase 3, respectively, that precipitated with PKCζ. The asterisk indicates mouse anti-FLAG immunoglobulin G light chains which react with the secondary antibody used to detect mouse anti-caspase 3. (B) Time course of the effect of 0.7 M NaCl on the interaction between PKCζ and caspase 9 and the effect of mutation of Ser144 to Ala (S/A).