Fig. 3: EGFR-KDD forms inter-molecular dimers and higher order oligomers after ligand stimulation.
a YAMC cells were cultured in serum-free medium for 12 h and then treated with 50 ng/mL EGF ligand for 5 min. Total EGFR and the autophosphorylation at three tyrosine sites were assessed by western blot. b YAMC cells were starved for 12 h and treated with cetuximab (10 µg/ml in serum-free medium) for 3 h 45 min, and EGF ligand (50 ng/mL in serum-free medium) was added for 15 min. The cells were harvested and analyzed by Western blot. WT, EGFR-WT; KDD, EGFR-KDD. c Template-based structural models of the intracellular portion of the EGFR-KDD inter-molecular dimer based on end-to-end EGFR-WT tetramer models. d Template-based structural models of EGFR-KDD inter-molecular dimer based on side-by-side EGFR-WT tetramer model. e Cross-correlation values of EGFR-WT and EGFR-KDD with (+) or without (−) ligand (EGF) stimulation is shown. The blue box indicates the ƒc value region for dimers. For the box and whiskers plot, the whiskers show the maximum and the minimum; the box shows 25th–75th percentile; and the line in the box is the median value. The median values are reported next to the boxplot. Each gray dot represents the averaged acquisition (10 s, 6 acquisitions) per area per cell. All data points are shown. Numbers in parenthesis above the boxplot are the total number of cells that data were taken on. For a and b, n = 3 experiments were repeated independently with similar results. Results are the representative of three independent experiments. For e, one-way ANOVA test with uncorrected Fisher’s LSD post hoc test were performed to obtain adjusted and individual P values. Data are presented as median values ± SD. For a, b and e, source data are provided in the Source Data file.
