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. 2011 May 26;301(2):G338–G346. doi: 10.1152/ajpgi.00057.2011

Fig. 2.

Fig. 2.

Heparin-binding EGF-like growth factor (HB-EGF) ligand binding mediates ErbB4 transactivation by TNF. A: YAMC-ErbB4 cells were incubated with either control mouse IgG or an ErbB4 blocking antibody for 1 h before the addition of TNF (100 ng/ml) for 30 min or HRG (10 ng/ml) for 10 min. B: YAMC-ErbB4 cells were incubated with an ErbB4 blocking antibody for 1 h before the addition of 10 ng/ml HRG, 1 ng/ml betacellin (BTC), or 1 ng/ml EGF for 10 min. Phosphorylated ErbB4 (PY-1284), EGFR (PY-1068), ErbB2 (PY-1248), ErbB3 (1289), or total actin was assessed by Western blot. YAMC-ErbB4 cells were incubated for 1 h with HB-EGF neutralizing antibody (C), HRG neutralizing antibody (D), or BTC neutralizing antibody (E) before stimulation with 100 ng/ml TNF for 30 min and with 50 ng/ml HB-EGF, 10 ng/ml HRG, or 10 ng/ml BTC for 10 min. Treatment with mouse IgG served as the negative control. Cell lysates were analyzed by Western blot with antibodies specific for phosphorylated ErbB4 (PY-1284), total ErbB4, or actin. All blots are representative of at least 3 experiments.