I have treated myocytes with a HDACi ( phenylbutyrate, 5 mM, 48h) and prepared RIPA cell lysates. In WB, together with acetylated, total total histones are also increasing. The levels are basal in control cells. I presume total histone levels should remain without change following HDACi treatment and only acetylated forms should change. Any idea about this trend?
How many times you run the gel? Is it reproducible? Try 3-5 times with different set of cell lysates to run the gel. I also expect that the total histone should not change.
I would recommend you to repeat the experiments and use another housekeeping gene as a loading control. It would be interesting if this is true. But I would suspect not. You can read this paper from Cell in 2013,
Article Acetylation-Mediated Proteasomal Degradation of Core Histone...
Thanks all.
I did repeat the expt and quantitated histones in total cell lysates and nuclear protein extract. In both ways found levels increasing with treatment. WI'll read the paper and get back.
thank you all.
@ Ashwin: will look for cell cycle markers to assess any prolongation in S-phase, hope it solves the mystery
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