supplementary Fig. 1 Anti-HSP70 Ab Anti-SHP2 Ab Anti-SHP2 Ab (Input) IP : HSP70 Ab IP : FLAG (SHP2) Ab Anti-SHP2 Ab Anti-HSP70 Ab Anti-HSP 70 Ab (Input) supplementary Fig. 1 FLAG SHP2 D.M.(EGF) FLAG SHP2 D.M FLAG SHP2 W.T.(EGF) FLAG SHP2 W.T. con Sup Fig 1. Lysates from cells that were transfected transiently with the FSHP2WT or the FSHP2DM constructs with or without EGF (100ng/ml) treatment for 10 min. The cell lysates were immunoprecipitated with either FLAG or HSP70 antibodies, followed by western blot analysis using SHP2 or HSP70 antibodies.

supplementary Fig. 2 Anti-4G10 Ab Anti-HSP70 Ab Anti-SHP2 Ab EGF 5 15 SHP2 DM/HSP70 SHP2 WT/HSP70 (cell lysates) EGF 5 15 (min) (kDa) 175 83 62 48 33 25 (100ng/ml) supplementary Fig. 2 Sup Fig 2. Western blot analysis was carried out with lysates of the cells transfected with SHP2WT or SHP2DM with HSP70 constructs. Indicated antibodies were used to detect specific proteins.

supplementary Fig. 3A supplementary Fig. 3B FLAG(GAB1) HSP70 SHP2 GAB1 + – SHP2WT FLAGGAB1 Transfection EGF (10 min) Input (100ng/ml) supplementary Fig. 3A + – HSP70 SHP2WT FLAGGAB1 Transfection EGF (10 min) GAB1 SHP2 IP : SHP2 Ab (first lane with rabbit IgG) IP : FLAG Ab (first lane with mouse IgG) Phos-GAB1 PI-3 kinase (100ng/ml) supplementary Fig. 3B Sup Fig 3A. Lysates were obtained from cells that had been transfected transiently with the FGAB1, the SHP2WT/FGAB1 or the SHP2WT/FGAB1/HSP70 constructs. The cell lysates were western-blotted with FLAG, HSP70, or SHP2 antibodies. Sup Fig 3B. The binding level of SHP2 with GAB1, HSP70 or other tyrosine phosphorylated protein was assessed by IP with SHP2 and FLAG antibodies. After IP, the indicated antibodies were used for western blot.