American Society of Hematology

Figure 4

$Figure 4. Orai1 protein abundance in sgk1−/− and sgk1+/+ platelets and megakaryocytes. (A) Confocal microscopy of Orai1 protein abundance in platelets from sgk1+/+ (left panels) and sgk1−/− (right panels) mice. Red represents actin; and green, Orai1. Bar represents 10 μm. (B) Flow cytometric analysis of Orai1 surface expression in platelets from sgk1+/+ (black bar) and sgk1−/− (gray bar) mice. (Bottom) Arithmetic mean ± SEM (n = 7). **P < .01. (Top) Representative overlays of Orai1-positive sgk1+/+ (black line) and sgk1−/− (gray line) platelets; light gray panel represents isotype control. (C) Western blot analysis of whole cell lysate protein of Orai1 and STIM1 from sgk1+/+ and sgk1−/− platelets. Arithmetic mean ± SEM (n = 4) of Orai1 protein abundance in sgk1+/+ (black bar) and sgk1−/− (gray bar) platelets. **P < .01. (D) Western blot analysis of isolated membrane protein and cytosolic fraction of Orai1 from sgk1+/+ and sgk1−/− platelets. Arithmetic mean ± SEM (n = 4) of Orai1 protein abundance in sgk1+/+ (black bars) and sgk1−/− (gray bars) platelets. **P < .01. (E) Confocal microscopy of Orai1 protein abundance in megakaryocytes cultivated from bone marrow of sgk1+/+ (left panels) and sgk1−/− (right panels) mice (top). Red represents GPIb; green, Orai1; and blue, nuclei. Bar represents 10 μm. Statistical analysis of Orai1 immunofluorescence abundance (bottom). **P < .01. n = 4.$

Orai1 protein abundance in sgk1^−/− and sgk1^+/+ platelets and megakaryocytes. (A) Confocal microscopy of Orai1 protein abundance in platelets from sgk1^+/+ (left panels) and sgk1^−/− (right panels) mice. Red represents actin; and green, Orai1. Bar represents 10 μm. (B) Flow cytometric analysis of Orai1 surface expression in platelets from sgk1^+/+ (black bar) and sgk1^−/− (gray bar) mice. (Bottom) Arithmetic mean ± SEM (n = 7). **P < .01. (Top) Representative overlays of Orai1-positive sgk1^+/+ (black line) and sgk1^−/− (gray line) platelets; light gray panel represents isotype control. (C) Western blot analysis of whole cell lysate protein of Orai1 and STIM1 from sgk1^+/+ and sgk1^−/− platelets. Arithmetic mean ± SEM (n = 4) of Orai1 protein abundance in sgk1^+/+ (black bar) and sgk1^−/− (gray bar) platelets. **P < .01. (D) Western blot analysis of isolated membrane protein and cytosolic fraction of Orai1 from sgk1^+/+ and sgk1^−/− platelets. Arithmetic mean ± SEM (n = 4) of Orai1 protein abundance in sgk1^+/+ (black bars) and sgk1^−/− (gray bars) platelets. **P < .01. (E) Confocal microscopy of Orai1 protein abundance in megakaryocytes cultivated from bone marrow of sgk1^+/+ (left panels) and sgk1^−/− (right panels) mice (top). Red represents GPIb; green, Orai1; and blue, nuclei. Bar represents 10 μm. Statistical analysis of Orai1 immunofluorescence abundance (bottom). **P < .01. n = 4.

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