Anti htt mw8

Paraffin-embedded liver sections, 40 μm per section, from male 16-wk-old R6/1 mice containing the Cre and floxed alleles of IKKβ (HD), treated with tamoxifin (n = 5) or oil vehicle control (n = 4), were used. One section per mouse was analyzed. Fluorescent aggregate immunolabeling was done with anti-HTT MW8 (AB528297; Developmental Studies Hybridoma Bank) (recommendation from Jeffrey Carroll for liver HTT, Western Washington University, Bellingham, WA) and TO-PRO-3 Iodide stain (T3605; Thermo Fisher Scientific) was used to label nuclei. Images were captured using Leica confocal microscope (DM2500) and Leica camera TCS SPE (Leica Microsystems Inc.). All images were taken at 20×, two areas per section, with consistent settings for laser intensity, exposure time, and gain through all images. In each area, the number of aggregates was counted by eye without normalization, as aggregate numbers were very low and visible per field.

The following antibodies and dilutions were used in this study: Anti-HTT Ab1 (aa1–17,(Difiglia et al., 1995 (link))) 1:50 for capillary immunoassay and 1:2000 for western blot; anti-HTT EPR5526 (Abcam, Waltham, MA, ab109115, 1:2000 for western blot); anti-polyQ MW1 (MilliporeSigma, Burlington, MA, MABN2427, 1:50 for capillary immunoassay); anti-polyQ PHP3 (generous gift from Dr. Ali Khoshnan, 1:2000 for western blot); Anti-PDE10A (Abcam, Waltham, MA, #ab177933, 1:2000 for western blot); Anti-DARPP32 (Abcam, #ab40801, 1:2000 for capillary immunoassay); Anti-GFAP (MilliporeSigma, Burlington, MA, AB5804, 1:3000 for capillary immunoassay); Anti-GAPDH (MilliporeSigma, Burlington, MA, #MAB374, 1:10000 for western blot); Anti-Sodium channel subunit beta-4 (Abcam, Waltham, MA, #ab80539, 1:500 for western blot); Anti-vinculin (Sigma, St. Louis, MO, #V9131, 1:5000 for capillary immunoassay, 1:2000 for western blot); Anti-ATP5A (Abcam, Waltham, MA, #ab14748, 1:2000 for western blot); Anti-HTT MW8 (University of Iowa Developmental Studies Hybridoma Bank, 1:1000 for filter trap); Anti-HTT S830 (generous gift from Dr. Gillian Bates, (Landles et al., 2010 (link)) 1:8000), HDAC1 (Abcam, Waltham, MA, ab32369–7, 1:4000).