Figure 1. Diffusion of various cargo... | Rockefeller University Press

Figure 1.

$Figure 1. Diffusion of various cargo proteins in the ER was severely restricted by hypertonicity. (A) The indicated areas of a COS7 cell expressing VSVG-YFP at 40°C were photobleached as described in Materials and methods. Images of a whole cell before and after photobleaching are shown. After the first series of FRAP (top), the osmolarity was adjusted to 930 mOsm, and images were recorded at 1, 30, and 60 min. The pink arrowhead indicates a spot photobleached at 1 min, and the yellow arrowhead indicates a spot photobleached at 30 min. (B) Quantitative FRAP of various cargo proteins when exposed to hypertonicity. Cells expressing misfolded VSVG-YFP (40°C), folded VSVG-YFP (32°C) in the presence of 10 μM brefeldin A, NHK, or tyrosinase (40°C) were exposed to 930 mOsm. FRAP was recorded before (black) and 1 (red) or 60 min (green) after treatment. The normalized mean recovery rates are plotted. The number of experiments and mobile fractions are shown in Table I. Bars, 5 μm.$

Diffusion of various cargo proteins in the ER was severely restricted by hypertonicity. (A) The indicated areas of a COS7 cell expressing VSVG-YFP at 40°C were photobleached as described in Materials and methods. Images of a whole cell before and after photobleaching are shown. After the first series of FRAP (top), the osmolarity was adjusted to 930 mOsm, and images were recorded at 1, 30, and 60 min. The pink arrowhead indicates a spot photobleached at 1 min, and the yellow arrowhead indicates a spot photobleached at 30 min. (B) Quantitative FRAP of various cargo proteins when exposed to hypertonicity. Cells expressing misfolded VSVG-YFP (40°C), folded VSVG-YFP (32°C) in the presence of 10 μM brefeldin A, NHK, or tyrosinase (40°C) were exposed to 930 mOsm. FRAP was recorded before (black) and 1 (red) or 60 min (green) after treatment. The normalized mean recovery rates are plotted. The number of experiments and mobile fractions are shown in Table I. Bars, 5 μm.

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