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. 2010 Jul 14;24(9):1805–1821. doi: 10.1210/me.2010-0071

Figure 2.

Figure 2

More of HA-MC4R-GFP bearing obesity-linked mutations than of wt receptor is ubiquitinated. A and B, N2A cells were transiently transfected with Flag-ubiquitin (Flag-ub) or cotransfected with Flag-ubiquitin and wt-HA-MC4R-GFP or mutated HA-MC4R-GFP (P78L, R165W, I316S, and I317T). Cell lysates were analyzed by Western blot (WB) by using the indicated antibodies. C and D, Immunoprecipitations (IP) from cell lysates were carried out with anti-HA antibodies and analyzed by Western blot by using the indicated antibodies. Arrows indicate the bands at 250 and 78 kDa. Vertical bar indicate MC4R immunoreactivity between 78 and 250 kDa. *, Unspecific bands.