American Society of Hematology

Figure 5

$Figure 5. PTPROt overexpression inhibits SYK Y352 tyrosyl phosphorylation. Tet-inducible Flag-tagged WT or mutant (CS) PTPROt clones were cultured with or without Dox and stimulated with goat anti–human IgG for 5 minutes (A), 15 minutes (B), or left untreated. Thereafter, cells were lysed and immunoprecipitated with anti-SYK or anti-CD79a antibody. Immunoprecipitates were size fractionated, blotted, and analyzed with the indicated phosphotyrosine antibodies (anti-pSYK352, -pSYK525/526, -pSYK323, and 4G10, respectively). The blots was subsequently stripped and blotted with an anti-pan SYK or anti-CD79a antibody. In each experiment, the corresponding whole cell lysates were simultaneously size-fractionated, blotted, and analyzed with Flag antibody to confirm Dox-induced overexpression of WT-PTPROt or CS-PTPROt.$

PTPROt overexpression inhibits SYK Y352 tyrosyl phosphorylation. Tet-inducible Flag-tagged WT or mutant (CS) PTPROt clones were cultured with or without Dox and stimulated with goat anti–human IgG for 5 minutes (A), 15 minutes (B), or left untreated. Thereafter, cells were lysed and immunoprecipitated with anti-SYK or anti-CD79a antibody. Immunoprecipitates were size fractionated, blotted, and analyzed with the indicated phosphotyrosine antibodies (anti-pSYK352, -pSYK525/526, -pSYK323, and 4G10, respectively). The blots was subsequently stripped and blotted with an anti-pan SYK or anti-CD79a antibody. In each experiment, the corresponding whole cell lysates were simultaneously size-fractionated, blotted, and analyzed with Flag antibody to confirm Dox-induced overexpression of WT-PTPROt or CS-PTPROt.

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