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. 2021 Apr 28;296:100720. doi: 10.1016/j.jbc.2021.100720

Figure 1.

Figure 1

Phosphorylation of Y311 on PKCδ is agonist dependent.A, schematic showing the locations of several tyrosine phosphorylation sites on PKCδ. Mouse and human sites are listed. An “Hu” prior to the number denotes human. B, western blots showing Y311 phosphorylation of PKCδ following stimulation of platelets with 500 μM AYPGKF with or without 10 μM Indomethacin, 100 μM MRS2179 and 500 nM AR-C69931MX. The combination of indomethacin, MRS2179 and AR-C69931MX are referred to as feedback inhibitors (FBIs). Washed human platelets were stimulated with 500 mM AYPGKF for the indicated time and the resulting platelet proteins were separated via SDS-PAGE. C, quantification of fluorescent band intensity from “B” expressed a ratio of pY311 to total PKCδ. D, western blots showing Y311 phosphorylation of PKCδ following stimulation (3 min) of human platelets with 5 μg/ml CRP with or without 100 nM YM-254890 and 500 nM AR-C69931MX. YM-254890 and AR-C69931MX are FBIs in this experiment. E, quantification of “D” expressed as a ratio of pY311 to total PKCδ, which was used to assess loading. ∗p < 0.05, n = 5. CRP, collagen-related peptide; PKC, protein kinase C.