Fig. 2: Impact of AMPK hyper-activation on synaptic markers expression.

Primary neurons were treated at 20 DIV for the indicated times with the AMPK activator, AICAR (1 mM). a Cell lysates were analyzed by WB with antibodies directed against phospho-AMPK-Thr172 (pAMPK), AMPK, phospho-ACC-Ser79 (pACC), ACC, and actin. b, c Quantification of the ratios pAMPK/AMPK (b) and pACC/ACC (c). Results represent mean ± SD, n = 6. d Cytotoxicity was assessed using the lactate dehydrogenase (LDH) assay after 24, 48, 72, and 96 h of AICAR treatment. Treatment with 0.9% Triton X-100 was used as a positive control. Results represent mean ± SD, n = 6. e–m Synaptic markers expression upon AMPK hyper-activation with AICAR treatment. WB analysis (e) and quantification of the pre-synaptic markers SNAP25 (f), Munc (g), synapsin Ia (Syn Ia) (h) and IIb (Syn IIb) (i), and the post-synaptic markers PSD-95 (j), Homer 1bc (k), GluN1 (l), and GluA1 (m). Results represent mean ± SD, n = 6. n–r Post-synaptic densities extractions were performed after 48 h AICAR treatment. Resulting non PSD and PSD fraction were analyzed by WB (n) and expression of the post-synaptic markers PSD-95 (o), Homer 1bc (p), GluN1 (q), and GluA1 (r) was quantified in the PSD fraction. Results represent mean ± SD, n = 4. One-way ANOVA with Bonferroni’s post hoc test was used for b–d and f–m experiments and Student’s t-test was used for o–r experiments; *p < 0.05, **p < 0.01, ***p < 0.001 compared to Ctrl