Fig. 4: EGFR-KDD directly interacts with ERBB family members.
a V5-epitope-tagged EGFR-WT and EGFR-KDD was co-transfected with Myc-epitope-tagged EGFR-WT and EGFR-KDD in HEK293 cells. After 48 h transfection, cells were lysed by hypotonic buffer and the cell lysates were immunoprecipitated by using V5 antibody. Immunoblotting were probed by V5 and Myc antibody. b Cross-correlation values of co-transfected EGFR-WT (mCherry-fused) and EGFR-KDD mutant (eGFP-fused) with (+) or without (−) ligand (EGF) stimulation is shown. The light blue box indicates the ƒc value region for dimers. c Myc-epitope-tagged EGFR-KDD was co-transfected with V5-epitope-tagged EGFR-WT, HER2, and HER3 in HEK293 cells. Cell lysates were immunoprecipitated by using V5 antibody. Immunoblotting were probed by V5 and Myc antibody. d Cross-correlation values of co-transfected HER2 (mCherry-fused) and EGFR-KDD mutant (eGFP-fused) with (+) or without (−) ligand (EGF) stimulation is shown. e Cross-correlation values of co-transfected HER3 (mCherry-fused) and EGFR-KDD mutant (eGFP-fused) with (+) or without (−) ligand (EGF) stimulation is shown. For a and c, n = 2 experiments were repeated independently with similar results. Results are the representative of two independent experiments. For b, d and e, data are reported as 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 where data were taken on. 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–e, source data are provided as a Source Data file.
