Fig. 1: Mutations disrupting the potential intra-molecular dimer interface abrogate phosphorylation of EGFR-KDD and anchorage-independent growth. | Nature Communications

Fig. 1: Mutations disrupting the potential intra-molecular dimer interface abrogate phosphorylation of EGFR-KDD and anchorage-independent growth.

From: Structure–function analysis of oncogenic EGFR Kinase Domain Duplication reveals insights into activation and a potential approach for therapeutic targeting

Fig. 1

a Ribbon diagram and space-filling model of EGFR-KDD kinase domains. Mutations constructed in this study were labeled. b Schematic representation of mutations we constructed in this study. We generated point mutations disrupting the potential intra- (C1, N2) and inter-molecular (N1, C2) dimer interface as well as mutations inactivating kinase activity of each kinase domain (Dead1, Dead2). c YAMC cells stably expressing EGFR-KDD and its mutants. Cells were cultured for 48 h and then harvested and lysed for analysis. Total EGFR and auto-phosphorylation at three tyrosine sites were evaluated by western blot. n = 3 experiments were repeated independently with similar results. EV empty vector; WT, EGFR-WT; KDD, EGFR-KDD. d Soft agar assays were performed in six-well plates by using YAMC cells. 5000 cells were seeded in each well and colonies were counted after 4 weeks. n = 3 biologically independent replicates were examined over three independent experiments with similar results. Data are presented as mean values ± SD. One-Way ANOVA test with Bonferroni post hoc test was performed to obtain the adjusted P values. For a, the model coordinates are provided in Supplementary Data 2. For c and d, results are the representative of three independent experiments. Source data are provided as a Source Data file.

Back to article page