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. 2011 Jun 30;25(9):1650–1660. doi: 10.1210/me.2011-1020

Fig. 4.

Fig. 4.

Hsc70 increases cell surface levels of intracellular retained MC4R mutants and coimmunoprecipitates (coIP) with the receptor. A, HEK293 cells were transfected with vectors for expression of WT or mutant MC4R and Hsc70-V5, or V5 (empty vector). Cell surface levels of mutant MC4R are shown relative to WT protein. Error bars represent 2× se. *, P < 0.05. B–E, Confocal analysis of HEK293 cells expressing WT (B), S58C (C), P78L (D), or D90N (E) HA-MC4R in combination with GFP or Hsc70-GFP. Arrow indicates cell surface expression. Scale bar, 10 μm. F, HEK293A cells were transfected with plasmids for expression of HA-MC4R alone or in combination with Hsc70-V5. Cells were lysed, and immunoprecipitation (IP) of HA-MC4R was then performed. IP were IB with anti-HA and anti-V5. Hsc70-V5 is indicated by an arrow, and an asterisk highlights immunoglobulin.