Figure 1
Figure 1. CEACAM1 is expressed on the surface and in intracellular pools in murine and human platelets. (A) Flow cytometric analysis of CEACAM1 surface and total expression on resting murine platelets. Platelets were stained with a polyclonal anti-murine CEACAM1 2457 antibody followed by a secondary FITC-conjugated anti-rabbit antibody. Normal rabbit serum and ceacam1−/− platelets were included as negative controls. Data were collected through a live platelet gate based on forward versus side scatter profiles on a FACSCanto flow cytometer (BD Biosciences). Results combine data derived from 3 independent experiments and are represented as mean fluorescence intensity (MFI) plus or minus SEM. (B) Flow cytometric analysis of CEACAM1 surface and total expression on resting human platelets. Platelets were stained with a monoclonal anti–human CEACAM1 4D1C2 antibody followed by a secondary FITC-conjugated anti–mouse antibody. Normal mouse IgG and ceacam1−/− platelets were included as negative controls. Data were collected through a live platelet gate based on forward- versus side-scatter profiles on a FACSCanto flow cytometer. Results combine data derived from 3 independent experiments and are represented as MFI plus or minus SEM. (C,D) Representative histogram profiles of CEACAM1 surface and total expression on resting murine and human platelets. CEACAM1 surface and total expression was determined as described in panels A and B. (E) Agonist stimulation of murine platelets using thrombin (0.25-1.00 U/mL) over a dose-dependent range. CEACAM1 surface expression was determined as described in panel A. (F) Agonist stimulation of human platelets using thrombin (0.25-1.00 U/mL) over a dose-dependent range. CEACAM1 surface expression was determined as described in panel B. (G) Flow cytometric analysis of CEACAM1 surface expression on resting murine platelets from wild-type versus ceacam1+/− and ceacam1−/− mice. CEACAM1 surface expression was determined as described in panel A. NRS indicates normal rabbit serum. (H) Representative histogram profiles of CEACAM1 surface expression on resting murine platelets from wild-type versus ceacam1+/− and ceacam1−/− mice. *P < .05; **P < .005.

CEACAM1 is expressed on the surface and in intracellular pools in murine and human platelets. (A) Flow cytometric analysis of CEACAM1 surface and total expression on resting murine platelets. Platelets were stained with a polyclonal anti-murine CEACAM1 2457 antibody followed by a secondary FITC-conjugated anti-rabbit antibody. Normal rabbit serum and ceacam1−/− platelets were included as negative controls. Data were collected through a live platelet gate based on forward versus side scatter profiles on a FACSCanto flow cytometer (BD Biosciences). Results combine data derived from 3 independent experiments and are represented as mean fluorescence intensity (MFI) plus or minus SEM. (B) Flow cytometric analysis of CEACAM1 surface and total expression on resting human platelets. Platelets were stained with a monoclonal anti–human CEACAM1 4D1C2 antibody followed by a secondary FITC-conjugated anti–mouse antibody. Normal mouse IgG and ceacam1−/− platelets were included as negative controls. Data were collected through a live platelet gate based on forward- versus side-scatter profiles on a FACSCanto flow cytometer. Results combine data derived from 3 independent experiments and are represented as MFI plus or minus SEM. (C,D) Representative histogram profiles of CEACAM1 surface and total expression on resting murine and human platelets. CEACAM1 surface and total expression was determined as described in panels A and B. (E) Agonist stimulation of murine platelets using thrombin (0.25-1.00 U/mL) over a dose-dependent range. CEACAM1 surface expression was determined as described in panel A. (F) Agonist stimulation of human platelets using thrombin (0.25-1.00 U/mL) over a dose-dependent range. CEACAM1 surface expression was determined as described in panel B. (G) Flow cytometric analysis of CEACAM1 surface expression on resting murine platelets from wild-type versus ceacam1+/− and ceacam1−/− mice. CEACAM1 surface expression was determined as described in panel A. NRS indicates normal rabbit serum. (H) Representative histogram profiles of CEACAM1 surface expression on resting murine platelets from wild-type versus ceacam1+/− and ceacam1−/− mice. *P < .05; **P < .005.

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