Fig 1: SRC, YES1, and FYN (SYF) kinases phosphorylate SHP2 Y62, Y542, and Y580.(A) Kinase Library prediction heatmap based on the peptide primary sequence of the SHP2 Y62 phosphosite plotted on human kinome evolutionary tree. (B) SFK multi-kinase and selective inhibitors with targets shown, tested here. (C-D) Immunoblot analysis of (C) U-2 OS cells and (D) HCC827 cells treated with dasatinib and SU6656 at indicated concentrations, and DMSO. (E-F) Immunoblot analysis of HCC827 cells treated with (E) saracatinib, CH6953755, and PP2, and (F) double and triple combinations at indicated concentrations, and DMSO. (G) Immunoblot analysis of SYF knock out and wildtype MEFs. (H) In vitro kinase assays with recombinant SRC, YES1, and FYN kinases and recombinant SHP2 WT and Y62F. The SHP2 pY542 antibody demonstrated nonspecific binding in the control conditions.
Fig 2: SRC, YES1, and FYN (SYF) kinases phosphorylate SHP2 Y62, Y542, and Y580.(A) Kinase Library prediction heatmap based on the peptide primary sequence of the SHP2 Y62 phosphosite plotted on human kinome evolutionary tree. (B) SFK multi-kinase and selective inhibitors with targets shown, tested here. (C-D) Immunoblot analysis of (C) U-2 OS cells and (D) HCC827 cells treated with dasatinib and SU6656 at indicated concentrations, and DMSO. (E-F) Immunoblot analysis of HCC827 cells treated with (E) saracatinib, CH6953755, and PP2, and (F) double and triple combinations at indicated concentrations, and DMSO. (G) Immunoblot analysis of SYF knock out and wildtype MEFs. (H) In vitro kinase assays with recombinant SRC, YES1, and FYN kinases and recombinant SHP2 WT and Y62F. The SHP2 pY542 antibody demonstrated nonspecific binding in the control conditions.
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