Cells were seeded so that they would reach at ~90% confluency on the following day. After one wash with PBS, cells were treated with the medium supplemented with the vehicle solvent, sodium propionate (Sigma-Aldrich, P5436), sodium valproate (Sigma-Aldrich, P4543), SAHA (Sigma-Aldrich, SML0061), β-hydroxybutyrate (Sigma-Aldrich, H6501), TSA (Sigma-Aldrich, T8552), sodium butyrate (Sigma-Aldrich, B5887), or HDAC3 inhibitor RGFP966 (Selleckchem, S7229) for 24 hours. Cells were washed with PBS and lysed in the RIPA buffer (Thermo Fisher Scientific, 89900) to prepare soluble protein extracts or in the Triton extraction buffer (PBS containing 0.5% Triton X-100, 2 mM phenylmethylsulfonyl fluoride, 0.02% NaN3, and protease inhibitors) to prepare protein extracts. Extracts were prepared in buffer K and further analyzed by immunoblotting with various antibodies as described above.

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