Figure 2

Role of AMPK in cannabinoid-induced autophagy. Western blot analysis of (a) LC3-II after treatment of Panc1 cells for 12 h with 200 μM GW or ACPA and 20 μM CC, and of (b) p-p70S6K and (c) LC3-II after treatment of Panc1 cells for 12 h with 200 μM GW or ACPA, in the presence or absence of the expression vectors for the AMPK wt or mutant R531G gamma-2 subunit. The bands were scanned as digital peaks and the areas of the peaks were calculated in arbitrary units, as described in Materials and Methods. The value of α-tubulin was used as a normalizing factor. Values are the means of three independent experiments (±S.D.). (d) AMP/ATP ratio was measured after 12 h treatment with 200 μM GW or ACPA, in the absence or presence of a pre-treatment for 1 h with 20 mM NAC, as reported in Materials and Methods. Values are the means of three independent experiments (±S.D.). Statistical analysis: *P<0.05, **P<0.01, and ***P<0.001