18 research outputs found

    The difference in cytosolic esterase-induced fluorescence with CMFDA between resting and activated human platelets.

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    <p>The activity of cytosolic esterase-induced fluorescence reaction is visualized with CMFDA. The fluorescence of resting platelets was clearly visible in laser confocal microscopy, whereas activated platelets release no or little fluorescence. A, C: Resting platelets; B, D: Activated platelets (activated by 100μM ADP); A, B: green fluorescence; C, D: bright field.</p

    CD62P expression percentage on membrane of platelets during 7-day storage at 22± 2°C.

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    <p>A: Mouse IgG1 κ Iso control. B: platelets stored for 1 day. C: platelets stored for 3 days. D: platelets stored for 5 days. E: platelets stored for 7 days.</p

    PS externalization of platelets during 7-day storage at 22 ± 2°C.

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    <p>A: platelets stored for 1 day. B: platelets stored for 3 days. C: platelets stored for 5 days. D: platelets stored for 7 days.</p

    Correlation of platelet CEIFI with functional parameters of platelets during 7-day storage.

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    <p>A: Correlation to the ADP-induced aggregation activity, r = 0.9813. B: Correlation to the HSR activity, r = 0.9848. C: Correlation to the expression percentage of CD62P on platelet membrane, r = -0.9945. D: Correlation to the PS externalization percentage, r = -0.9847.</p

    The change of cytosolic esterase-induced fluorescence activity in platelets during 7-day storage at 22 ± 2°C.

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    <p>The activity of cytosolic esterase-induced fluorescence in platelets is measured with CMFDA. *<i>P</i> <0.05 as compared with platelets stored for 1 day.</p

    HSR curves of platelets during 7-day storage at 22± 2°C.

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    <p>A: platelets stored for 1 day. B: platelets stored for 3 days. C: platelets stored for 5 days. D: platelets stored for 7 days.</p

    Aggregation curves of platelets during 7-day storage at 22 ± 2°C.

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    <p>A: platelets stored for 1 day. B: platelets stored for 3 days. C: platelets stored for 5 days. D: platelets stored for 7 days.</p

    PAEP gene knockdown in melanoma cells.

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    <p>Stable PAEP 624.38-Mel shRNA transfectant and its non-targeting negative control transfectant, shControl, were established with a corresponding shRNA lentivirus. PAEP gene expressions were assayed by semi-quantitative RT-PCR (A) and Western blotting (B). PAEP protein secreted by melanoma cells was quantitated by ELISA (C).</p

    Activation of T helper lymphocytes by melanoma-derived PAEP protein.

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    <p>(A-B) The percent of CD3<sup>+</sup> T cells in unfractionated PBLs from healthy donors was assayed for by direct staining with anti-CD3-FITC followed by flow cytometric analysis. (A) Negative control, (B) Healthy donor. Flow cytometry was applied to confirm the purity of CD4<sup>+</sup> T cells before (C) and after (D) MACS isolation. (E) Levels of cytokines secreted by PHA-stimulated T helper cells co-cultured with PAEP-rich 624.38-Mel shControl or PAEP-poor 624.38-Mel shPAEP supernatant were determined. Melanoma-derived PAEP significantly inhibited both IL-2 and IFN- γ secretion by T helper cells (* p<0.05, Student’s <i>t</i>-test). (F) Levels of cytokines secreted by PHA- or anti-CD3 antibody-stimulated Th1 cells co-cultured with PAEP-rich 624.38-Mel shControl or PAEP-poor 624.38-Mel shPAEP supernatant were determined. Melanoma-derived PAEP significantly inhibited both IL-2 and IFN-γ secretion by Th1 cells (* p<0.05, Student’s <i>t</i>-test) (G-H) There were no differences in number of CD4/CD8 cells before or after PHA stimulation in the presence of PAEP (1 μg/ml). (G) PBLs; (H) CD3<sup>+</sup> T cells. (I-J) The number of CD69/CD44 cells before and after activation in the absence or presence of PAEP was determined by direct staining with antibodies and flow cytometry. The number of CD69 cells significantly decreased co-cultured with PAEP-rich 624.38-Mel shControl supernatant. The experiment was repeated at least three times.</p
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