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. Author manuscript; available in PMC: 2019 Apr 26.
Published in final edited form as: FASEB J. 2007 Jul 11;21(14):4095–4100. doi: 10.1096/fj.06-7927com

Figure 3.

Figure 3

Subcellular fractionation of the MC2R-Nup50 complex. H295R cells were fractionated as described [27] into four fractions (whole cell lysates, membrane, crude nuclear, and purified nuclear fractions). (a) Demonstrates the contents of subcellular fraction for an ER marker (GRP78/BiP) and nuclear membranes (Nup62) by immunoblotting. (Fraction 1, whole cell lysate; fraction 2, membranes; fraction 3, crude nuclear extract; fraction 4, purified nuclear extract). (b) In membrane fractions the MC2R-Nup50 complex is demonstrated by co-immunoprecipitation using the MC2R C-16 antibody at rest and following stimulation by ACTH (10−6M) for the initial 30 minutes. Some reappearance of the complex occurs by 150 minutes. (c) in purified nuclear fractions the MC2R-Nup50 complex is not detectable at rest, but begins to appear by 60 minutes and is prominent at 150 and 210 minutes.