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Blood, Vol. 110, Issue 9, 3183-3191, November 1, 2007 This Article Abstract Full Text Services Email this article to a friend Alert me to new issues of the journal Rights and Permissions Citing Articles Citing Articles via CrossRef Megakaryocyte-restricted MYH9 inactivation dramatically affects hemostasis while preserving platelet aggregation and secretion Blood Léon et al. 110: 3183 Supplemental materials for: Leon et al Files in this Data Supplement: Document 1. Supplemental methods (PDF, 94.2 KB) Figure S1. Generation of a platelet-specific MYH9Δ knock-down strain (PDF, 77.4 KB) - (A) LoxP sites (▲) were inserted from part to part of exon 1 from MYH9 (i, ii). The construct was linearized and electroporated into ES cells. Homologous recombinant clones were screened by Southern Blot (probe shown in (i) and by PCR amplification (open arrows) and then transiently transfected with a plasmid expressing Cre recombinase to delete the neomycin selection cassette and to obtain the floxed MYH9Flx allele (iii). Cells with the floxed allele were injected into blastocysts to produce chimeric mice. The deletion of the MYH9 allele (MYH9) was obtained after the floxed allele was exposed to in vivo cre recombinase expression (iv). (B) Southern blot analysis using the probe shown in (A,(i)). The wt allele is 6.6 kb in length and the floxed allele is 8.3 kb. (C) PCR analysis of wt, heterozygous or homozygous MYH9Flx using primers apart from the first LoxP site (small arrows). Video 1. Intravital observation of thrombus growth following FeCl3-induced injury of the carotid artery of wt mice after busulfan-induced thrombocytopenia (WMV, 4.69 MB) - Platelets were labeled with DIOC6 and the growth of the thrombus was recorded based on fluorescence intensity. Images were acquired sequentially (1 image/s) and the movie was accelerated 10 times. Video 2. Intravital observation of thrombus growth following FeCl3-induced injury of the carotid artery of MYH9Δ mice (WMV, 4.69 MB) - Platelets were labeled with DIOC6 and the growth of the thrombus was recorded based on fluorescence intensity. Images were acquired sequentially (1 image/s) and the movie was accelerated 10 times. This Article Abstract Full Text Services Email this article to a friend Alert me to new issues of the journal Rights and Permissions Citing Articles Citing Articles via CrossRef

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