Acetylation of Cytidine Regulates Translation in Prefrontal Cortices of MK-801-exposed C57BL/6 Mice

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original data for western blot.

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Total proteins were extracted from the dissected tissue in 500 μL RIPA lysis buffer [50 mM Tris pH 7.4, 150 mM NaCl, 1% Triton, 0.5% sodium deoxycholate, 0.5% SDS, 5 mM EGTA, protease inhibitor cocktail (Roche)]. After homogenization and centrifugation at 16,200× g for 10 min, protein in the supernatants was quantified using the BCA protein assay, and the samples were diluted with a 6× SDS loading buffer (125 mM Tris, pH 6.8, 1% SDS, 300 mM DTT, 30% glycerol, and 0.01% bromophenol blue). Protein samples were separated by SDS-PAGE in 10% polyacrylamide gels, transferred to PVDF membranes (Millipore), and immunoblotted. Primary antibodies were diluted in 0.1% TBS-Tween as follows: mouse anti-β-Actin (Proteintech Cat# 66009-1-Ig, 1:2500), rabbit anti-NAT10 (Abcam Cat# ab194297, 1:1000), rabbit anti-NMDAR2A (Abcam Cat# ab124913, 1:1000).

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