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Blood, Vol. 113, Issue 3, 675-678, January 15, 2009
R93W mutation in Orai1 causes impaired calcium influx in platelets
Blood Bergmeier et al.
113: 675
Supplemental materials for: Bergmeier et al
Files in this Data Supplement:
- Document 1. Supplemental materials and methods (PDF, 104 KB)
- Figure S1. Phenotypic characterization of Orai1R93W chimeras (JPG, 30.9 KB)
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Isolated leukocytes from two Orai1R93W chimeras as well as CD45.1 and CD45.2 control mice were stained for surface expression of CD45.1 antigen and B cell marker B220. B cells were gated by B220 positivity. FITC indicates fluorescein isothiocyanate. It is important to note that B cells from both chimeras were CD45.1-negative. Since recipient mice were CD45.1-positive while transplanted fetal liver cells were CD45.1-negative, all the B cells (and likely all other blood cells) in the two chimeras shown here derived from the donor fetal liver cells.
- Figure S2. Normal expression of key adhesion receptors on the surface of Orai1R93W platelets (JPG, 22.1 KB)
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WT and Orai1R93W platelets were stained with fluorophore-labeled antibodies to the β1 integrin subunit, CD9, GPIbα, and GPIX, and analyzed immediately by flow cytometry. n=5.
- Figure S3. Semi-quantitative detection of Orai1, Orai2, and Orai3 RNA in human platelets and megakaryocytic cells (JPG, 23 KB)
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RNA for Orai paralogues was detected in purified platelets and Meg-01 cells by RT-PCR. CD41 was used as a control platelet marker. “No RNA,” control using cDNA reaction that contained no RNA input. All PCR products are of the expected size.
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