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. Author manuscript; available in PMC: 2021 Jul 7.
Published in final edited form as: Mol Neurobiol. 2019 Jun 25;56(12):8345–8363. doi: 10.1007/s12035-019-01659-w

Fig. 6. NRG2 shedding by PMA requires NMDAR activity and the ADAM10 selective blocker GI254023X blocks NMDAR-mediated NRG2 shedding.

Fig. 6

(a) Representative Western blot of whole cell lysates from transduced hippocampal neurons showing the effects of PMA (0.2 μM, 10 min), without or with 100 μM AP5 pre-treatment, on pro-NRG2 protein levels and electrophoretic mobility. AP5 and NMDA treatments are included as controls. (b) Corresponding ELISA measurements of ecto-NRG2 in culture supernatants treated as described in (a). Data are normalized to AP5 control and represent the mean ± SEM of 6 data points from 3 independent experiments. *, p<0.05; n.s., not significant (F(3,20) = 36.54; 1-way ANOVA with Tukey’s multiple comparison). (c) Representative Western blot of whole cell lysates from transduced hippocampal neurons showing inhibition of NMDAR-dependent pro-NRG2 processing in cultures that were pre-incubated with GM6001 (10 μM) or GI254023X (3 μM). (d) Corresponding ELISA measurements of ecto-NRG2 in culture supernatants treated as described in (d). Data are normalized to AP5 controls and represent the mean ± SEM of 6 data points from 3 independent experiments. n.s., not significant (F(3,20) = 85.27; 1-way ANOVA with Tukey’s multiple comparison).