Computational protocol: Class IIa Histone Deacetylases Are Conserved Regulators of Circadian Function*

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Protocol publication

[…] To assess the effect of HDAC inhibitors on BMAL1 acetylation, NIH3T3 cells transfected with BMAL1-Myc using FuGENE 6 (Roche Applied Science) were treated with indicated HDAC inhibitors 24 h after transfection. After 18 h, cells were lysed in hot SDS sample buffer. To assess the effect of HDAC5 expression on BMAL1 acetylation protein extracts were prepared from cells 40 h after transfection. Proteins were resolved by SDS-PAGE using pre-cast Bis-Tris gels (Invitrogen) and immunoblotted using standard protocols. Band intensities were quantified with ImageJ software. Statistical significance was determined with a two-tailed paired Student's t test using the SPSS Statistical suite (IBM). […]

Pipeline specifications

Software tools ImageJ, SPSS
Applications Miscellaneous, SPIM, Microscopic phenotype analysis
Organisms Mus musculus, Drosophila melanogaster, Homo sapiens