Fig. 2

HDAC4 is endogenously expressed in the mushroom body and has similar distribution to the HDAC4^WT transgene. A Immunohistochemistry with anti-GFP on HDAC4::EGFP brains. Top row. The left-most image is a Z-stack maximum projection through the anterior of the brain showing the pattern of endogenous HDAC4 expression. The images to the right are 1-μm sections through the α, β and γ lobes of the mushroom body showing that HDAC4 is present in each of the lobes. Scale bar = 100 μm. Middle row. 0.5-μm optical sections through the Kenyon cell layer showing the distribution of HDAC4 in the calyx (asterisk) and cell bodies. Scale bar = 50 μm. Bottom row. Magnification of the images directly above, showing the cytoplasmic haloes around the nuclei (counterstained with DAPI), and punctate foci within some nuclei (arrows). Scale bar = 20 μm. B Immunohistochemistry with anti-GFP and anti-Myc on whole mount HDAC4::EGFP brains expressing HDAC4^WT-Myc driven by OK107-GAL4. Note that constitutive OK107 drives much stronger expression than maximal induction with GAL80ts; therefore, the aggregates are larger than in Fig. 1D. HDAC4^WT-Myc is visible in in cytoplasmic haloes and in aggregates in a subset of nuclei. Within these nuclei, endogenous HDAC4 is also pulled into the aggregates. C Western blot of endogenous HDAC4 in the absence and presence of HDAC4^WT transgene expression. Anti-HDAC4 detects products ~ 135 kDa consistent with the estimated size of HDAC4 isoforms. When expressed with elav-GAL4, HDAC4^WT was detected at slightly larger molecular weight (red arrow) due to the 6xMyc tag, and the identity of HDAC4^WT was confirmed via detection of the Myc tag