Fig. 1.

Screening of ENT-A011. ENT-A011 induces TrkB and its downstream target Akt phosphorylation after 20 min treatment in NIH-3T3 TrkB stable expressed cells. Representative blots (A) and quantification A’ of 8 (TrkB) and 6 (Akt) independent experiments are shown. Error bars represent S.E.M., Student’s t-test against Control; *p < 0.05; ***p < 0.001. B, B′ ENT-A011 protects NIH-3T3 TrkB cells from cell death caused by serum deprivation. Representative images (B) and quantification of Toxicity Assay (B′) on NIH-3T3 TrkB cells after treatment with BDNF or compound ENT-A011 for 24 h with or without TrkB inhibitor, ANA-12. (without ANA-12: BDNF N = 10/ENT-A011 N = 10, with ANA-12: BDNF N = 7/ENT-A011 N = 9), error bars represent S.E.M., Student’s t-test against Control; *p < 0.05, **p < 0.01, ***p < 0.001. C, C′ Nomad biosensor activation via TrkB phosphorylation in HEK 293 TrkB transiently transfected cells after ENT-A011 treatment. Receptor activation leads to increase in fluorescence levels. Representative images (C) and dose response curve for different compound concentrations (C′) are shown. Scalebars = 100 μm