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. Author manuscript; available in PMC: 2019 Jan 18.
Published in final edited form as: Mol Cell. 2018 Jan 18;69(2):279–291.e5. doi: 10.1016/j.molcel.2017.12.024

Figure 1. AMPK Suppresses EZH2-mediated Histone H3K27 Trimethylation.

Figure 1

(A) Immunoblot (IB) analysis of whole cell lysates (WCL) derived from Ampkα WT and Ampkα1−/−; Ampkα2−/− double knock out (DKO) MEFs.

(B) Ampkα1−/−; Ampkα2−/− DKO MEFs were infected with the retroviral construct expressing HA-AMPKα1. Infected cells were selected with 1 μg/ml puromycin for 72 hours to eliminate the non-infected cells before harvesting.

(C) Ampkα WT and DKO MEFs were treated with 100 μM A769662 for the indicated period of time before harvesting.

(D) T98G cells were treated with 2 mM metformin for 2 days before harvesting.

(E) Ampkα WT and DKO MEFs were infected with shGFP control or shEzh2 lentiviral shRNA. The infected cells were selected with 1 μg/ml puromycin for 72 hours to eliminate the non-infected cells before harvesting.

(F) Quantification of the relative H3K27me3 band intensities from three independent experiments. H3K27me3 bands were normalized to TUBULIN, and then normalized to the first lane. Data are represented as mean ± SD, n=3. * P < 0.05, Student’s t test.

(G) shGFP- (as a negative control) and shEZH2-MDA-MB-231 cells were treated with or without 10 μM AMPK inhibitor Compound C for 12 hours before harvesting for IB analysis.

See also Figure S1.