HDAC3 preferably binds to nuclear Htt with long Qs.

<p><b>A:</b> GST-HDAC3 binds directly to either cytoplasmic or nuclear Htt-ex1. GST pull-down assay of E1, E2, E3 (cytoplasm), and N1, N2, N3 (nuclear) HeLa cell lysates is shown. Pulled-down fraction was analyzed by anti-GFP or GST antibodies. *Non-specific band. <b>B:</b> HDAC3 immunoprecipitates almost exclusively with nuclear Htt-ex1s. E1, E3 (cytoplasm), N1, and N3 (nuclear) HeLa cells were transfected with FLAG-tagged HDAC3 and those lysates were immunoprecipitated with anti-FLAG antibodies immobilized to protein G agarose beads. The pre-IP fraction and the IPed fraction were analyzed using anti-FLAG or anti-GFP antibodies. Molecular weight markers are shown on the left. <b>C:</b> HDAC3 associates exclusively with nuclear inclusion bodies. E3 or N3 cells were fixed and stained with anti-HDAC3 antibodies and visualized by Alexa 546 conjugated secondary antibodies. Arrowheads: inclusion bodies with no HDAC3 signals associated. Arrows: HDAC3 signal-associated inclusion bodies. Bar = 20 µm.