American Society of Hematology

Fig. 4.

$Fig. 4. Immunoprecipitation of Mst1 and Mst2 from apoptotic eosinophils. / (A) Eosinophil lysates were obtained from freshly purified eosinophils and from cells cultured for 24 hours in cytokine-free medium. Following immunoprecipitation with antibodies to the N-terminal catalytic fragments of Mst1 and Mst2, the pellet (P) and supernatant (S) fractions were separated on 10% gels by SDS-PAGE and the kinase activities measured by “in gel” renaturation assays using MBP as a substrate. (B) Supernatant obtained following initial immunoprecipitation (R1) was immunoprecipitated for a second (R2) and third (R3) time using the antibody to Mst1 and the activity in the pellets determined by MBP “in gel” renaturation assay. Representative autoradiographs are presented and are typical of 3 independent experiments.$

Immunoprecipitation of Mst1 and Mst2 from apoptotic eosinophils.

(A) Eosinophil lysates were obtained from freshly purified eosinophils and from cells cultured for 24 hours in cytokine-free medium. Following immunoprecipitation with antibodies to the N-terminal catalytic fragments of Mst1 and Mst2, the pellet (P) and supernatant (S) fractions were separated on 10% gels by SDS-PAGE and the kinase activities measured by “in gel” renaturation assays using MBP as a substrate. (B) Supernatant obtained following initial immunoprecipitation (R1) was immunoprecipitated for a second (R2) and third (R3) time using the antibody to Mst1 and the activity in the pellets determined by MBP “in gel” renaturation assay. Representative autoradiographs are presented and are typical of 3 independent experiments.

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