Figure 4
TrkB-ErbB4 interaction was increased following NRG1 treatment. (A) Immunoprecipitation (IP) with the anti-TrkB antibody or control IgG antibody followed by blotting with the anti-ErbB4 or anti-TrkB antibody was performed in neurons (DIV4) after NRG1 stimulation with or without K252a pretreatment. (B) IP with the anti-ErbB4 antibody or control IgG antibody followed by blotting with the anti-TrkB or anti-ErbB4 antibody. (C) Inhibition of NRG1-induced increase in TrkB-ErbB4 interaction in neurons (DIV4) with a BDNF neutralizing antibody. The upper panel shows a representative autoradiogram of TrkB and ErbB4, and the lower panel represents normalized TrkB values. Data represent mean ± SEM. (n = 3). *P < 0.05 vs control; $P < 0.05 vs NRG1 treatment. (Di) PLA was performed in DIV4 cortical neurons following treatment with NRG1 for 5 min. The antibodies were omitted in the control PLA group (control PLA). The images were acquired at 20X or 40X and scale bar is 20 μM. The red spots represent the interaction between TrkB and ErbB4. (Dii) Quantitation of PLA signal was performed in three independent experiments using imageJ. Data represents mean PLA signal/cell ± SEM. *P < 0.05 vs control.