Fig. 4
From: Cathelicidins prime platelets to mediate arterial thrombosis and tissue inflammation
Cathelicidin-dependent signaling in platelets. a–i LL-37 induced signaling in isolated human platelets. a–c Flow cytometry analysis of platelet P-selectin surface expression in the presence of a the calcium chelator BAPTA (n = 5) or a phospholipase C inhibitor (U-73122, n = 6), b pertussis or cholera toxin to inhibit G-protein signaling (n = 4), or c inhibitors of tyrosine kinases Src-family kinases (Dasatinib, n = 7) and Syk (R406, n = 5). d Representative western blots of phosphorylated Src-family kinase and phosphorylated Syk upon incubation of platelets with LL-37. Collagen was used as positive control for tyrosine kinase phosphorylation, β-actin served as loading control. Images are representative of three independent blots. e–i Flow cytometry analysis of LL-37 platelet P-selectin surface expression in the presence of e STAT3 small molecule inhibitor (Stattic, n = 6), f GPVI antibody (HGP5C4, n = 9), g GPIIb/IIIa antibody (Abciximab or Tirofiban, n = 4), h formyl-peptid-receptor (FPR1 or FPR2) antibody, and i inhibitors against the purinergic P2X7-receptor (Boc-MLF 10, WRW4 or A438079, n = 4). j–l CRAMP-induced signaling in isolated mouse platelets. j P-selectin surface expression in the presence of a GPVI depleting antibody (JAQ1, n = 6). k P-selectin surface expression and l phospholipase C phosphorylation in platelet lysates from platelet-specific Syk-deficient mice and respective littermates after stimulation with CRAMP (flow cytometry: n = 10 Syk−/− and n = 5 Syk+/+ animals, western blot analysis n = 5 each). Graphs show mean and SEM. P-values were determined by unpaired (a, f, j–l) or paired (c, e, h) t-test
