Figure 2.

Mandelalide A induces time and concentration-dependent phosphorylation of AMPK and ACC that is suppressed by the AMPK inhibitor dorsomorphin. (A) Immunoblot analysis of MEFs treated with mandelalide A (30 nM or 100 nM) for 30 min or 90 min. Whole cell lysates were probed with antibodies for phospho-AMPK, phospho-ACC, total AMPK, total ACC and GAPDH as indicated. Blots are representative of a single experiment that was repeated three times. (B) Histograms show the quantification of immunoblot data shown in A from three independent experiments; values represent band intensity of phospho-AMPK/total AMPK and phospho-ACC/total ACC, normalized to GAPDH. (C) Immunoblot analysis of MEFs treated with mandelalide A (30 nM) or oligomycin A (1 µM) with, or without, dorsomorphin (10 mM) for 1 h. Whole cell lysates were probed with antibodies as indicated. Blots are representative of a single experiment that was repeated at least three times. (D) Histograms show quantification of immunoblot data shown in C; values represent band intensity of the phosphorylated protein, relative to total (p-AMPK/total AMPK and p-ACC/total ACC) normalized to GAPDH. Statistical significance of change relative to vehicle (0.1% DMSO) is indicated in B and D as * p < 0.05, ** p < 0.01.