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. 2019 Nov 19;20(22):5817. doi: 10.3390/ijms20225817

Figure 1.

Figure 1

HER family analysis. (A) EGFR phosphorylation antibody array revealed phosphorylation of HER2 (T686 and S1113), HER3 (T1289), and HER4 (T1284), as well expression of EGFR, HER2, HER3, and HER4. One milligram of total protein lysate was used for IP experiments, and 20 µl of immunoprecipitated protein was used for WB. A431 (EGFR/HER3) and SKBr3 (HER2) cell lysates were used as positive control for p- and total protein. (B) EGFR IP/WB experiments confirmed phosphorylation (upper box) and expression (bottom box) of EGFR. (C) HER2 IP/WB experiments confirmed phosphorylation (upper box) and expression (middle box) of HER2; HER2/EGFR co-IP experiments revealed HER2-EGFR heterodimerization (bottom box). (D) HER3 IP/WB experiments confirmed phosphorylation (upper box) and expression (bottom box) of HER3. (E) EGFR/HER3 co-IP experiments revealed HER3-EGFR heterodimerization. (F) HER3/HER2 co-IP experiments revealed HER3-HER2 heterodimerization; HER3 WB revealed HER3 expression. (G) HER3 (red) and EGFR (green) IF provided evidence of HER3 and EGFR co-expression (yellow). (H) IHC showed HER3 immunoreactivity. Legend: IP: immunoprecipitation; WB: Western blotting; IF: immunofluorescence; IHC: immunohistochemistry.