Figure 2
Both sc-tPA and tc-tPA at 300 nM can modulate EGFR signaling. (a) Representative immunoblots for phospho-EGFR (A–C: tyrosine 1173; D–F: tyrosine 992) and total EGFR on neurons (12–13 DIV) after treatments with EGF (50 ng/ml), sc-tPA or tc-tPA (300 nM) during 15 min. (B, C, E and F) Quantifications of phosphorylated EGFRs and total EGFRs were compared with control (mean±S.E.M.; N=3 or 4 experiments; *P<0.05). (g) Confocal images of endogenous NMDAR–EGFR complexes in cortical neurons. (H) Quantification of NMDAR–EGFR complexes detected by Proximity Ligation Assay (PLA). ***P-value<0.001; Mann–Whitney U-test, N=3. (I) Cross-immunoprecipitation assays demonstrating that EGFRs and NMDARs form stable complexes in cultured neurons. (J) After two NMDA stimulations used as baseline, neurons were incubated for 45 min with buffer (control, n=85 cells) or EGF 50 ng/ml (EGF, n=93 cells) prior to a second set of NMDA stimulations. Percentages of potentiation or inhibition after incubation are calculated for each cell. (K) Percentage of potentiation or inhibition after incubation for each group (mean±S.E.M.; *P<0.0001 Kruskal–Wallis test followed by Mann–Whitney U-test; #P<0.0001 Wilcoxon test comparison of preincubation and postincubation responses)
