Syk^{R41Afl/fl;PF4-Cre} signaling is inhibited in response to GPVI and CLEC-2 agonists. WT and Syk^{R41Afl/fl;PF4-Cre} platelets (R41A) (5 × 10⁸/mL) were stimulated with either rhodocytin (300 nM for 300 seconds) (A,C) or CRP (3 μg/mL for 90 seconds) (B) followed by lysis. Where stated, platelets were preincubated with either PP2 (20 μM) or PRT318 (5 μM). Proteins were subsequently immunoprecipitated (IP). For the time course (D), platelets were stimulated with rhodocytin (300 nM), and samples were removed after 1, 3, and 5 minutes to prepare whole-cell lysates. Precipitated proteins and whole-cell lysates were then separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and western blot (WB) for phosphotyrosine (n = 3). Blots were exposed with either autoradiographic film (A-B) or using a Licor Odyssey FC (C-D). Quantitation was performed with Licor Image Studio and normalized to percentage of the WT/maximal response. Where absent, there was no difference compared with background.