Journal: Nature medicine

Article Title: PPARδ repression in Huntington’s disease and its essential role in CNS translate into a potent agonist therapy

doi: 10.1038/nm.4003

Figure Lengend Snippet: (a) 3X-PPRE luciferase reporter activity in primary cortical neurons from BAC-HD or non-transgenic (Non-Tg) control mice, co-transfected with Renilla luciferase vector, and PPARδ and/or PGC-1α expression constructs, as indicated, and treated with GW501516 (100 nM) or vehicle. Results were normalized to Non-Tg neurons at baseline. * P < .05; t-test. (b) Mitochondrial membrane potential of primary cortical neurons from BAC-HD and Non-Tg mice, treated as indicated, was determined from the ratio of mitochondrial to cytosolic JC-1 fluorescence. Results were normalized to Non-Tg neurons at baseline, and CCCP treatment served as a positive control for depolarization. * P < .05, ** P < .01; t-test. (c) Immunofluorescence of active caspase-3 (red) and microtubule-associated protein 2 (green) in primary cortical neurons from BAC-HD and Non-Tg mice, treated as indicated. Scale bar = 20 μm. (d) Quantification of active caspase-3 staining shown in (e) . Results were normalized to Non-Tg neurons at baseline. * P < .05, ** P < .01; t-test. (e) Cortex from Non-Tg mice immunostained with the indicated PPAR antibody (green) and NeuN antibody (red). Merged images reveal expression of indicated PPAR. Scale bar = 50 μm. (f) Immunofluorescence of active caspase-3 in primary cortical neurons from BAC-HD mice, transfected with the indicated shRNA construct for 3 days, or treated with fenofibrate 100 nM (PPARα agonist), GW501516 100 nM (PPARδ agonist), or pioglitazone 20 nM (PPARγ agonist) for 24 h, prior to exposure to 25 μM H 2 O 2 . * P < .05, ** P < .01; ANOVA with post-hoc Tukey. Error bars = s.e.m. All experiments were performed with 3 biological replicates and 9 technical replicates per condition.

Article Snippet: Addgene provided constructs for PPARα (22751) and PPARγ (8895).

Techniques: Luciferase, Activity Assay, Transgenic Assay, Control, Transfection, Plasmid Preparation, Expressing, Construct, Membrane, Fluorescence, Positive Control, Immunofluorescence, Staining, shRNA