FIGURE 3.
Lyn C-terminal inhibitory tyrosine residue (pY507) is a direct substrate of SHP2. (A) WT and KO BMMCs were starved of IL-3 for 6 h and stimulated with SCF (50 ng/ml) for indicated times. Lysates were subjected to IB analysis with pY507-Lyn, pYSrc 416 (cross-reacts with pY396 Lyn) and Lyn Abs. (B and C) Line graph depicting the relative phosphotyrosine levels of Lyn at tyrosines 507 and 396, respectively, at indicated time points (mean ± SD; n = 3; *p < 0.05, significant difference between genotypes). (D) Lysates were prepared from SCF-treated Baf/3-KIT SHP2 KD cells (50 ng/ml for 5 min) and incubated with either GST or GST-PTPD/A:Q/A bound to beads overnight at 4°C. Mock reactions were performed with including lysates. IB analyses were carried out with Abs indicated on the right.