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Fig. 4 | Molecular Medicine

Fig. 4

From: Normal and Expanded Huntington’s Disease Gene Alleles Produce Distinguishable Proteins Due to Translation Across the CAG Repeat

Fig. 4

Huntingtin is located in the cytosolic fraction of lymphoblast cells

(A) Detection of huntingtin. A Western blot of proteins in subcellular fractions of lymphoblasts obtained from a normal individual (N) and an HD homozygote (HD) was probed with a 1:10,000 dilution of HF1. Fractions are denoted by PN, postnuclear supernatant; S, 100,000 × g supernatant (S100); P, 100,000 × g pellet (P100), and N, nucleosol. The position of the 200-kD marker (myosin) is shown, and the positions of the 350-kD normal (N) and HD (HD) huntingtin are indicated by arrows. The bands at 240 and 260 kD are also detected by the preimmune sera. (B) Detection of control proteins. Western blots of proteins in the subcellular fractions of lymphoblasts shown in Panel A were probed with control antibodies. The top panel shows the results obtained using a monoclonal antibody directed at pRB, a 110-kD nuclear protein that is detected in the N fraction (24), and the bottom panel the results with a monoclonal antibody directed at pERK1 (Transduction Laboratories), a 44-kD plasma membrane associated protein that is detected in the P fraction (25). The band at 45 kD is probably a phosphorylated form of ERK1 that is also recognized by this monoclonal. In separate experiments (not shown), similar Western blots were probed with polyclonal rabbit antisera against ornithine aminotransferase (OAT), a mitochondrial protein (26) that was detected in the P fraction and neurofibromin (27), a cytoplasmic protein that can associate with microtubules that was found in both the P and S fractions (28).

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