American Society of Hematology

Figure 1.

$Figure 1. SHIP-2 is expressed in a megakaryocyte cell line and in human platelets. (A) Membranes containing approximately 20 μg mRNA from the indicated cell lines, adenocarcinoma of the cervix (HeLa), megakaryoblast (MEG-01), T-cell leukemia (Jurkat), lung carcinoma (A549), neuroteratocarcinoma (NT,) and glioma (C6), were hybridized with SHIP-2 cDNA (nucleotides 3017-3989) and washed as described in “Materials and methods.” After exposure, the membrane was allowed to decay and was subsequently hybridized to an actin probe. (B) The Triton X-100– soluble fraction of platelets was isolated and immunoprecipitated with either nonimmune (Non I) sera, affinity-purified SHIP-2 antipeptide, or SHIP-1 antibodies and immunoblotted with antibodies to SHIP-2 (top), or SHIP-1 (bottom). (C) COS-1 cells transiently transfected with HA empty vector, or HA–SHIP-1 were harvested and the lysates immunoblotted with either SHIP-1 (left), or SHIP-2 (right) antibodies. (D) Platelets were either left untreated or treated with thrombin (1 U/mL, final concentration) for 5 minutes at room temperature and then lysed. The Triton-soluble and actin cytoskeletal (Actin CSK) fractions were isolated and immunoblotted with antibodies to SHIP-2, filamin, glycocalicin (GPIb), or p85 as indicated. Results shown are typical of 3 separate experiments.$

SHIP-2 is expressed in a megakaryocyte cell line and in human platelets. (A) Membranes containing approximately 20 μg mRNA from the indicated cell lines, adenocarcinoma of the cervix (HeLa), megakaryoblast (MEG-01), T-cell leukemia (Jurkat), lung carcinoma (A549), neuroteratocarcinoma (NT,) and glioma (C6), were hybridized with SHIP-2 cDNA (nucleotides 3017-3989) and washed as described in “Materials and methods.” After exposure, the membrane was allowed to decay and was subsequently hybridized to an actin probe. (B) The Triton X-100– soluble fraction of platelets was isolated and immunoprecipitated with either nonimmune (Non I) sera, affinity-purified SHIP-2 antipeptide, or SHIP-1 antibodies and immunoblotted with antibodies to SHIP-2 (top), or SHIP-1 (bottom). (C) COS-1 cells transiently transfected with HA empty vector, or HA–SHIP-1 were harvested and the lysates immunoblotted with either SHIP-1 (left), or SHIP-2 (right) antibodies. (D) Platelets were either left untreated or treated with thrombin (1 U/mL, final concentration) for 5 minutes at room temperature and then lysed. The Triton-soluble and actin cytoskeletal (Actin CSK) fractions were isolated and immunoblotted with antibodies to SHIP-2, filamin, glycocalicin (GPIb), or p85 as indicated. Results shown are typical of 3 separate experiments.

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