Fig. 3.

Katacine induces phosphorylation of CLEC-2 and key signaling protein on CLEC-2 signaling pathway. ( A ) Representative immunoblots of platelet lysates (4 × 10 ⁸ ) stimulated with DMSO 0.1% (negative control), rhodocytin at 100 nM (positive control) or katacine (10 µM). (A-i) Tyrosine phosphorylation profiles in platelets stimulated with katacine or rhodocytin have similar profile. (A-ii) An increase of the phosphorylation levels of Syk (Y525/526) and LAT (Y200) in platelets treated with rhodocytin 100 nM and katacine 10 µM is shown compared with the DMSO 0.1% control. (A-iii) The mean and SD of four independent experiments. ( B ) A representative blot of an immunoprecipitation of CLEC-2 from lysed platelets (4 × 10 ⁸ platelets/mL) and probed against the tyrosine phosphorylation antibody (4G10), demonstrating a significant increase on the CLEC-2 phosphorylation levels in platelets stimulated with 100 nM rhodocytin or 10 µM katacine compared with the DMSO 0.1% control. (B-i) The mean ± SD values from three independent experiments. The Kruskal–Wallis one-way ANOVA test was used to evaluate significant differences between the treatment in relation with the control; the symbol “*” represents significance difference where p  < 0.05, as stated. SD, standard deviation.