Figure 3
From: Cannabidiol induces autophagy via ERK1/2 activation in neural cells
CBD-induced autophagy by CB1, CB2 and TRPV1 receptor activation in neural cells. The autophagic flux was evaluated in SH-SY5Y and murine astrocyte cells after 2 h of treatment with 10 µM CBD and/or with pre-treatment (30 min) with the respective CB1, CB2 and TRPV1 antagonists, AM 251, AM 630 and capsazepine (CPZ) (10 µM each), in the presence or absence of the lysosome inhibitor NH4Cl, added during the last hour of treatment. CBD potentiated LC3-II levels under NH4Cl blockade, which was reverted by AM 251, AM 630 and CPZ pre-treatment in both SH-SY5Y (A-C) and murine astrocyte (D-F) cells. Samples were subjected to western blotting using anti-LC3 and anti-GAPDH antibodies. Representative images of LC3-II are shown in the panels above the bar graphs. The bar graph data is reported as the means ± S.E.M of LC3-II levels after GAPDH normalization. Analysis was conducted with Three-way ANOVA followed by Sidak’s post-hoc test; *p < 0.05, ** p < 0.01 and ***p < 0.001. The statistical summaries for CBD versus antagonist comparisons are detailed in Supplementary Tables S1 and S2. Entire blots are presented in Supplementary Figure S2.
