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. 2014 Jan 3;9(1):e84746. doi: 10.1371/journal.pone.0084746

Figure 4. Tyrosine phosphorylation of cellular substrates and formation of signaling complexes by Jak2-V617F and Lyn in PVTL-1 cells.

Figure 4

A–D) PVTL-1 cells were left untreated as control or treated with 1 µM JakI-1, 5 µM imatinib, or 100 nM dasatinib, as indicated for 6 h and lysed. Total cell lysates (A) and immunoprecipitates obtained with anti-Lyn (B), anti-Jak2 (C), or anti-CrkL (D) were subjected to Western blot analyses with antibodies against indicated proteins. Abbreviations used are: PY, phosphotyrosine; STAT5-PY, phospho-Y694-STAT5; Lyn-PY, phospho-Y396-Lyn; Jak2-PY, phospho-Y1007/1008-Jak2; TCL, total cell lysate; IP, immunoprecipitation. An asterisk indicates the position of the most significantly tyrosine-phosphorylated protein. Positions of tyrosine-phosphorylated and non-phosphorylated CrkL, CrkL-PY and CrkL-non-PY, respectively, are indicated. Positions of molecular weight markers are also indicated. E, F) PVTL-1 cells were left untreated as control or treated with 1 µM JakI-1or 100 nM dasatinib, as indicated for 6 h. Cell lysates were subjected to immunoprecipitation with anti-SHP1, anti-SHP2 (E), and anti-Gab2 (F) followed by Western blot analyses. Positions of relevant proteins are indicated.