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Image Search Results
Journal: JCI Insight
Article Title: Ubiquitin ligase Nedd4 regulates the abundance and toxicity of mutant huntingtin
doi: 10.1172/jci.insight.181013
Figure Lengend Snippet: ( A ) Nedd4 KD increases mHtt aggregation in primary cortical neurons. Mouse primary cortical neurons were transduced with lentivirus expressing Htt Exon1-25Q or -72Q together with scrambled (scr) or shNedd4-35 (N4-35). Cells were fixed and immunostained for Htt followed by confocal microscopy. DAPI was included in mounting medium to stain nuclei. ( B ) Nedd4 KD increases the size and the number of mHtt aggregates in primary cortical neurons. n = 5 microscopic fields for each, 2-tailed unpaired t test (*** P < 0.001). ( C ) Nedd4 KD enhances Htt toxicity in an mHtt expression-dependent manner in primary cortical neurons. Mouse primary cortical neurons were transduced with lentivirus expressing Htt Exon1-25Q or -72Q together with scrambled (scr) or shNedd4-35 (N4-35). Pictures were taken with bright-field microscope to show cell morphology.
Article Snippet: The following reagents were purchased as indicated: DMEM (11995065, Thermo Fisher), L-Glutamine (25030081, Thermo Fisher), Penicillin-Streptomycin (15140122, Thermo Fisher), Neurobasal media (21103049, Thermo Fisher), B-27 Supplement (17504044, Thermo Fisher), N-ethylmaleimde (23030, Thermo Fisher), PMSF (36978, Thermo Fisher), ZeptoMetrix RETROtek HIV-1 p24 Antigen ELISA (22-156-700, Fisher), FBS (100-106, Gemini Bio), CHX (14126, Cayman Chemical), bafilomycin A1 (11038, Cayman Chemical), bortezomib (#2204, Cell Signaling Technology), Ponseau S (P7170, Sigma), rapamycin (R8781, Sigma), cOmplete mini EDTA-free protease inhibitor cocktail (11836170001, Sigma), Protein A-Sepharose (P3391, Sigma), Protein G-Agarose (P4691, Sigma),
Techniques: Transduction, Expressing, Confocal Microscopy, Staining, Microscopy