Experiments were conducted in mouse C2C12 myoblasts and myotubes that were propagated in DMEM supplemented with 10% FBS. Cells were cultured to 80% confluence and passaged every 2–3 days at 37 °C in 5% CO2. C2C12 myoblasts can be differentiated into mature myotubes by replacing 10% FBS with 2% DES (horse serum) when cells reach 80% confluence. Differentiated myotubes were treated 4-days post differentiation as indicated in the results. Myoblasts and myotubes were pre-treated with HDAC inhibitors for 24 hrs prior to agonist(s) treatment. Myoblasts were pre-treated with siRNAs for 72 hrs prior to PMA treatment as indicated in figures and figure legends. All siRNAs were purchased from Sigma: nontargeting siRNA control (Cat#SIC001), siRNA HDAC1 (Cat# SASI_Mm01_0015_9497), siRNA HDAC1 (Cat#SASI_Mm01_00159502), siRNA HDAC2 (Cat#SASI_Mm01_0010_0070), siRNA HDAC2 (Cat#SASI_Mm01_00100_699), siRNA HDAC3 (Cat#SASI_Mm01_0016_2230) and siRNA HDAC8 (Cat#SASI_Mm01_0010_7278). Treatment times for phorbol-12-myristate-13-acetate (PMA), dexamethasone, or angiotensin II are indicated in figures and figure legends.
Habibian J.S., Bolino M, & Ferguson B.S. (2023). HDAC8 regulates protein kinase D phosphorylation in skeletal myoblasts in response to stress signaling. Biochemical and biophysical research communications, 650, 81-86.
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