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. Author manuscript; available in PMC: 2016 Feb 17.
Published in final edited form as: Circulation. 2014 Dec 17;131(7):656–668. doi: 10.1161/CIRCULATIONAHA.114.010186

Figure 4.

Figure 4

DUSP3-deficiency impairs Syk tyrosine phosphorylation. TLs were prepared from WT or Dusp3-KO mice platelets. Cells were non-activated, CRP- (0.3 μg/mL) or rhodocytin-activated (10 nM) for 30, 60, or 90 s. (A-C) Western blot analysis with anti-pY antibody (4G10) and with ERK1/2 (A) or GAPDH (C) as loading control. Arrows in (B) indicate unknown protein bands with attenuated pY levels in DUSP3-deficient platelets. (D-E) Representative pY blot of Syk immunoprecipitates from TLs of CRP- (D) or rhodocytin-activated (E) platelets. (F-G) Representative blot of Syk phosphorylation on Tyr-323 and Tyr-525/526 in CRP- (F) or rhodocytin-activated (G) platelets. Normalization was performed using total Syk. (H) pY and Syk western blots on FcRγ immunoprecipitates. (I-J) pY blot of PLCγ2 immunoprecipitates from TLs of CRP- (I) or rhodocytin-activated (J) platelets. Results shown are representative of 3 independent experiments performed on platelet pools from three mice each.