Figure 4
From: KRAS K104 modification affects the KRASG12D-GEF interaction and mediates cell growth and motility
Analysis of KRAS/GEF interactions. (A) Two branches of a thermodynamic cycle are placed in one simulation box. The different boxes in the scheme are indicated by the green (\(\lambda =0\)) and blue (\(\lambda =1\)) shading. The free energy estimate corresponds to a double free energy difference: ΔΔG = ΔG1 – ΔG2. The figure was generated by Microsoft PowerPoint 2016. (B) Interactions of KRAS with GEF and RASA1. KRAS was detected by western blot analysis after immunoprecipitation of endogenous GEF and RASA1 in H1299 cells after transfection with KRSWT, KRASG12D, KRASG12D/K104A or KRASG12D/K104Q plasmids. (C–F) Relative levels of GST-Raf1-RBD, RAS, Ac-lysine, p-AKT and AKT detected by western blotting after overexpression of KRASG12D, KRASG12D/K104A or KRASG12D/K104Q plasmids in (C,E) H1299 (KRASWT) and (D,F) MCAS (KRASMT) cancer cell lines. β-Actin was used as a loading control for cell lysates. The full-length and multiple exposures blots are presented in Supplementary Information.
