Endogenous expression of inactive lysine deacetylases KDAC4 (HDAC4) and KDAC7 (HDAC7)

To investigate the roles of KDAC4 and KDAC7, we generated cell lines containing point mutations in HT1080 cells that result in endogenous expression of mutated proteins. For KDAC4, we introduced a mutation (H803A) to inactivate catalysis but that also contains concurrent mutations resulting in a knockout, as well as one that enhances in vitro activity of KDAC4 (H976Y, heterozygous with a knockout mutation on the second chromosal copy). For KDAC7, an inactivating mutation (H709A, homozygous) was introduced. Expression profiling analysis was then performed on the resulting cell lines and wild-type HT1080 cells treated with the KDAC inhibitor TMP195, vs. untreated wild-type HT1080.