Fig. 6.

Intracellular retained mutant MC4R inclusion incidence is reduced and mobility at the ER increased in cells with elevated levels of Hsc70. A, Quantification of the incidence of inclusions in WT and mutant MC4R-GFP expressing cells with and without heterologous expression of Hsc70. A minimum of 50 transfected cells were counted for each treatment from three separate transfections. Counting was blind to experimental status 24 h after transfection. Error bars represent 2× se. B–D, Graphical representation of recovery after photobleaching of MC4R-GFP mutants S58C (B), P78L (C), and D90N (D) in cells cotransfected with a plasmid for expression of Hsc70-V5 or empty V5 plasmid (Con). A 2 × 2 μm area of the ER corresponding to the boxed areas was photobleached, and the cells images every second for 40 sec. Fluorescent intensities of the 2 × 2 μm area were normalized to prebleach levels at 100%. Error bars represent se, n = 15.