FIGURE 6.

Expression and activity of H141A mutant of HDAC1. A, a bar graph comparing deacetylase activity of the wild type and H141A mutant HDAC1-FLAG proteins expressed from plasmids transiently transfected into HDAC1 KO RD cells (clone #17) and immunoprecipitated with α-FLAG antibody before the assay. Negative control is untransfected HDAC1 KO RD cells. B, a work flow diagram for generation of HDAC1 transgene-expressing, WRN-depleted HDAC1 KO cells. Shown is the fastest timeline available. C, a Western blot measuring expression of wild type and H141A mutant HDAC1-FLAG within 10 days of transduction with pLenti-EFS-T2A-emGFP constructs in HDAC1 KO RD cells (clone #4). D, a Western blot measuring expression of WRN and of wild type and two isolates of H141A mutant HDAC1-FLAG after flow-sorting to purify HDAC1-expressing cells, their outgrowth, and siRNA transfection.