Differential expression of mutant NMMHC-IIA polypeptide in peripheral blood cells. Immunofluorescence analysis of peripheral blood smears from patients 1 and 2 double stained with PRB440P (red) and NT629 (green). (A) PRB440P stained granular NMMHC-IIA accumulation and diffusely stained cytoplasm of neutrophils, whereas NT629 stained only intense cytoplasmic foci and no background. (B) Lymphocytes were diffusely stained with both PRB440P and NT629. (C) In monocytes, both PRB440P and NT629 detected small dots in addition to diffuse cytoplasmic staining. Platelets in panels A-B are notably not stained with NT629. (D) PRB440P diffusely stained platelet cytoplasm and intensely stained cell periphery. In contrast, most platelets are NT629 negative; only extraordinarily large platelets are diffusely stained. (E) Mutant NMMHC-IIA polypeptide cannot translocate to lamellipodia in surface-activated platelets. Platelets were adhered to glass slides for 10 minutes, fixed, and processed for PRB440P and NT629 immunofluorescence analysis. Left column: images stained with May-Grünwald-Giemsa. Mutant NMMHC-IIA was weakly distributed only in granulomere zone of platelets from patients. Platelets from patients 2, 3, and 4 were examined. Original magnification, ×1000.