Figure 1.
ACTN1 p.Arg46Gln in the French macrothrombocytopenia pedigree with a model of autosomal dominant inheritance.
The arrow indicates the proband. The sequencing electropherogram of the heterozygous ACTN1 p.Arg46Gln missense mutation is shown in the right-hand bottom box.
Figure 2.
Morphology of platelet and MK cells from the proband and a control.
(A) Morphology of platelets in the bone marrow smears, which were stained with May-Grunwald Giemsa. The platelet in the proband is much larger than its normal size by reference to the red blood cells and to the control. The analysis was performed on an optical microscope (Leica DMR) with a 50×/1.40 numerical aperture oil objective lens (Leica). (B) Morphology and size (indicated by red arrows in the proband) of platelets from the peripheral blood observed by electronic microscopy (magnification of 10000×). (C) Morphology of a representative cultured MK from the bone marrow observed by electronic microscopy (magnification of 6000×). Ultrathin sections were examined by a JEM-1010 transmission electron microscope (JEOL) at an accelerating voltage of 80 kV.
Figure 3.
Evaluation of the functional effect of ACTN1 p.Arg46Gln on transfected COS-7 cells by immunofluorescence.
COS-7 cells transfected with either wild-type (top panel) or mutant (lower panel) ACTN1 plasmid construction (as well as non-transfected cells) were stained with three different flurochromes as indicated in the Figure. In the merged image, overexpressed α1-actinin, normal F-actin network and counterstained nuclei appear in green, red and blue, respectively. In the top right panel, the arrow indicates an untransfected cell. Fluorescence was visualized on an Olympus BX-61 fluorescence microscope with a 60×/1.40 numerical aperture oil objective lens (Plan Apo, Olympus). Image capture was performed with a Diagnostics Instruments CCD camera system using SPOT Advanced software version 3.2.4 (Diagnostic Instruments, Sterling Heights, MI). Image processing was made with Image G (1.45S, NIH, USA).