Fig. 3

NNI-362 putative mechanism via allosteric stimulation. Significant kinases that either are inhibited or stimulated by NNI-362 in CEREP panel of 151 kinases, significant with ± 25% inhibition (a). Measure of active site competition serial dilution to 10× (1000 nM) the effective concentration of NNI-362 (DiscoverX, KinomeScan) (b). Phosphorylation of p70S6 kinase in cultures prepared from 6 and 12DIV human neural progenitors treated with 0, 100, 300, and 1000 nM NNI-362 with each media change, n = 3 (c). Effect of NNI-362 on p70S6 kinase in two separate preparations of human neuronal progenitor cells at 5–6 and 11DIV (d). Phosphorylated kinases not consistently modulated by NNI-362: Akt1/2/3&pan, CREB, ERK1/2, GSK3beta, HSP 27, JNK1/2&pan, MKK3, MKK6, MSK2, p38alpha/beta/delta/gamma, p53, RSK1/2, and TOR. Effects of CDK5 inhibitors (indirubin and BML529), GSK3beta inhibitors (indirubin-3-monoxime and kenpaullone), and IKK2 inhibitor (SC-514) (e) on NNI-362-associated proliferation. Effects of p38 MAPK inhibitors on NNI-362-associated proliferation (f). Putative mechanism describing neuronal regeneration during aging and potential mechanism of action for NNI-362 (g). In vitro studies were analyzed using Kruskal-Wallis with group as factor. *p < 0.05 vs. vehicle, ^#p < 0.05 vs. NNI-362