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. 2014 Jan 9;57(3):642–650. doi: 10.1021/jm401837e

Figure 3.

Figure 3

Y23A, Y24A, R34A, and C151A mutants affect HDAC1 deacetylase activity but not protein association. (A) Wild-type or mutant proteins were expressed in T-Ag Jurkat cells as FLAG-tagged fusion proteins, immunoprecipitated with anti-FLAG-agarose beads, and tested for catalytic activity using an in vitro fluorescence assay (histogram) or separated on SDS-PAGE and probed with anti-FLAG antibody (gel image). The histogram shows the mean percent of at least four independent trials, which were normalized to wild-type deacetylase activity (set to 100%). The standard error is shown as error bars (Table S1). (B). Immunoprecipitates of wild type and alanine mutants in panel A were further probed with anti-FLAG, anti-RbAp48, or anti-mSin3A antibodies to assess protein association.