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. 2010 Jan 26;5(1):e8905. doi: 10.1371/journal.pone.0008905

Figure 2. gp78 interacts with htt.

Figure 2

A. gp78 interacts with Nhtt and polyglutamine expansion increases the interaction. Nhtt17Q and Nhtt138Q were expressed in HEK293 cells. FLAG-ATF6 and empty vector was used as controls. Immunoprecipitation was performed with anti-gp78 antibody followed by immunoblotting with the antibodies as indicated. B, C. gp78 enhances Nhtt association with the microsomes. HEK293 cells fractionated as indicated were subjected to fractionation into microsomal membranes (m) and cytosol (c). gp78 and Nhtt were detected by immunoblotting using anti-gp78 and anti-FLAG antibodies. Calnexin was blotted as an ER marker. Microsome-associated Nhtt was expressed as a percentage of total Nhtt associated with microsomes and in the cytosol (Mean ± SD, n = 3). D. Endogenous gp78 associates with htt. HEK293 cells were lysed in RIPA buffer and gp78 was immunoprecipitated with anti-gp78 antibody clone 2G5. Endogenous htt was detected by immunoblotting.