Fig. 3.

SH3BGRL enhances tyrosine site-specific HER2 phosphorylation. a Immunoblots of HER2 in SH3BGRL-overexpressing and knocking down cell lines. b Immunoblots of the indicated HER2 phosphorylation by SH3BGRL. SH3BGRL enhances EGF-induced HER2 tyrosine phosphorylation and prolongs the activation time, especially at pHER2 (Y1196) site. Cells were starved for 24 h, and stimulated with 20 ng/ml EGF for different time points (0, 5, 15 and 30 min). c Phosphorylation analysis of HER2 by western blotting assay. Transient co-transfection experiments were performed in HEK293T cells with YFP-HER2 and HA-SH3BGRL truncated mutants △α1, △α2, △β3. GAPDH was used as loading control. d Molecule docking prediction of the interaction between SH3BGRL and HER2. The model image was generated by Pymol. The upper panel shows the overview interaction of SH3BGRL (in purple) with C-terminal of HER2 (in blue). The lower panel presents the closest amino acid binding sites