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. 2020 May 18;117(22):12121–12130. doi: 10.1073/pnas.2000848117

Fig. 3.

Fig. 3.

Model PPI and MAFI by RAS–RBD interaction. (A) Diagrams depict the experimental design. The interaction of nl–KRAS4B_G12D and wild-type cl–RBD represents the conventional PPI. cl–RBD_R89L mutant serves as a negative control. The weak intrinsic affinity between nl–KRAS4B_G12D and membrane-associated cl–RBD_R89L–CVIM should mimic membrane association-facilitated interaction (MAFI). (B) ReBiL2.0 assay shows that KRAS4B_G12D interacts with RBD, but not RBD_R89L. The bar chart reports quantitative ReBiL2.0 scores normalized to the KRAS4B_G12D + RBD interaction (set to 100%). The 1/2luc alone lacking fusion partners (nl + cl) served as the negative control. Data show mean ± SEM (n = 4). (C) ReBiL2.0 assay shows that RBD_R89L–CVIM restores interaction with KRAS4B_G12D. The bar chart reports quantitative ReBiL2.0 scores normalized to the KRAS4B_G12D + RBD interaction (set to 100%). The 1/2luc alone lacking fusion partners (nl + cl) served as the negative control. Data show mean ± SEM (n = 4).