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Results: 11

1.
Figure 3

Figure 3. From: Insulin Action on GLUT4 Traffic Visualized in Single 3T3-L1 Adipocytes by Using Ultra-fast Microscopy.

Reversal of insulin action by wortmannin. Single 250-nm section through the middle of the cell taken before (C) and after 20 min of insulin stimulation (I). Wortmannin (+W) was added after 20 min of insulin and the cell reimaged 5 min later.

Varsha Patki, et al. Mol Biol Cell. 2001 January;12(1):129-141.
2.
Figure 11

Figure 11. From: Insulin Action on GLUT4 Traffic Visualized in Single 3T3-L1 Adipocytes by Using Ultra-fast Microscopy.

Effects of microtubule and actin depolymerization on insulin-stimulated 2-deoxyglucose uptake. Latrunculin-A (10 μM), nocodazole (10 μM), and cytochalasin-B (20 μM) were added 30 min before the addition of insulin (100 nM), as indicated along the abscissa. 2-Deoxyglucose uptake was measured after 15 min of insulin addition.

Varsha Patki, et al. Mol Biol Cell. 2001 January;12(1):129-141.
3.
Figure 8

Figure 8. From: Insulin Action on GLUT4 Traffic Visualized in Single 3T3-L1 Adipocytes by Using Ultra-fast Microscopy.

Effect of nocodazole on insulin-stimulated 2-deoxyglucose uptake. 3T3-L1 adipocytes were preincubated with buffer (no preincubation), with 10 μM nocodazole for 30 min (nocodazole, 30 min), or with 20 μM cytochalasin-B for 5 min. Insulin (100 nM) was then added, and after the times indicated on the abscissa, uptake of 2-[1,2-3H]deoxy-d-glucose was measured as described in MATERIALS AND METHODS.

Varsha Patki, et al. Mol Biol Cell. 2001 January;12(1):129-141.
4.
Figure 7

Figure 7. From: Insulin Action on GLUT4 Traffic Visualized in Single 3T3-L1 Adipocytes by Using Ultra-fast Microscopy.

Effect of nocodazole on GLUT4-eGFP dynamics. (A) 3T3-L1 adipocytes were incubated without (C) or with 10 μM nocodazole (N) for 20 min, fixed, and stained with antibody against alpha-tubulin followed by anti-mouse secondary antibody coupled with fluorescein isothiocyanate. (B) Single 250-nm optical sections through a 3T3-L1 adipocyte before (C) or after 20-min incubation with 5 μM nocodazole (N) (see QuickTime Demo 3).

Varsha Patki, et al. Mol Biol Cell. 2001 January;12(1):129-141.
5.
Figure 10

Figure 10. From: Insulin Action on GLUT4 Traffic Visualized in Single 3T3-L1 Adipocytes by Using Ultra-fast Microscopy.

Effect of latrunculin-A on GLUT4-eGFP and EEA1. 3T3-L1 adipocytes expressing GLUT4-eGFP (control) were exposed to Lat-A for 20 min, fixed, permeabilized, and stained with a polyclonal antibody raised to EEA1. Each panel is the projection in a single two-dimensional plane of 30 optical sections through the juxtanuclear region of the cell. The arrows on the top right panel point to vacuoles containing GLUT4-eGFP. Bar, 3 μm.

Varsha Patki, et al. Mol Biol Cell. 2001 January;12(1):129-141.
6.
Figure 9

Figure 9. From: Insulin Action on GLUT4 Traffic Visualized in Single 3T3-L1 Adipocytes by Using Ultra-fast Microscopy.

Effect of latrunculin-A on GLUT4-eGFP dynamics. (A) Single 250-nm optical sections through a 3T3-L1 adipocyte before (left) or after 20-min exposure to 10 μM Lat-A. The phase image (right) was obtained immediately after, to indicate that the cell remained spread out and attached to the coverslip. (B) 2D-projection of 21 optical sections through a 3T3-L1 adipocyte taken at 1-min intervals after 75 min of exposure to Lat-A. The arrow points to the trajectory of a single vesicle migrating toward the center of the cell (QuickTime Demos 4 and 5).

Varsha Patki, et al. Mol Biol Cell. 2001 January;12(1):129-141.
7.
Figure 6

Figure 6. From: Insulin Action on GLUT4 Traffic Visualized in Single 3T3-L1 Adipocytes by Using Ultra-fast Microscopy.

Motility of GLUT4-eGFP revealed by 2D projection of 4D images. 100 stacks of 21 sections taken before (CONTROL), or after insulin addition (INSULIN, 0 to 6.6 and INSULIN 6.6 to 13.2) were summed and displayed as 2D images. Arrows point to a region were a linear track is defined by the presence of a contiguous GLUT4-eGFP signal. Bottom panels are an amplification of the same region from the cell from the before and after insulin sets. Bars on the lower left corner represent 3 um.

Varsha Patki, et al. Mol Biol Cell. 2001 January;12(1):129-141.
8.
Figure 5

Figure 5. From: Insulin Action on GLUT4 Traffic Visualized in Single 3T3-L1 Adipocytes by Using Ultra-fast Microscopy.

Dynamics of GLUT4-eGFP in response to insulin. (A) Panels represent the 2D projection of 21-image sections taken every 4 s, as indicated in the upper left corner. Arrows point to structures near the juxtanucelar region, which display rapid changes in shape. The bar at the lower left corner of the first panel represents 1.7 μm. (B) Panels shown were taken after 5 min of insulin addition, at 4-s intervals, as indicated in the top left corner. The arrow follows the trajectory of a stream of GLUT4-eGFP from the juxtanuclear region to the plasma membrane (see QuickTime Demo 2).

Varsha Patki, et al. Mol Biol Cell. 2001 January;12(1):129-141.
9.
Figure 4

Figure 4. From: Insulin Action on GLUT4 Traffic Visualized in Single 3T3-L1 Adipocytes by Using Ultra-fast Microscopy.

Higher resolution imaging of GLUT4-eGFP. Images were acquired at settings that result in a resolution of 133 nm/pixel. The bar at the lower left corner of the first panel is 800 nm. (A) Panels represent a single 250-nm section through the middle of the cell acquired at the times (Min et al., 1999) after insulin addition indicated in the top left corner. Italicized numbers in the bottom right corner of each panel represent the percentage of maximal effect of insulin, which was calculated as described in Figure 3B. A phase image of the cell is shown in the first panel. (B) Panels represent a single 250-nm section through the bottom of the cell acquired at the times after insulin addition indicated in the top left corner.

Varsha Patki, et al. Mol Biol Cell. 2001 January;12(1):129-141.
10.
Figure 1

Figure 1. From: Insulin Action on GLUT4 Traffic Visualized in Single 3T3-L1 Adipocytes by Using Ultra-fast Microscopy.

Enhancement of resolution by image restoration. (Top) Four 250-nm optical sections of a 3T3-L1 adipocyte expressing GLUT4-eGFP. The number at the lower left corner of each image represents the image plane, the lower number being closest to the coverslip. (Middle) Same sections shown in the top, after restoration. (Bottom) Comparison of the information contained in a single 250-nm section from the middle of the cell (left) with the information contained in the 2D projection of 21 sections (middle). The large arrowhead in the expanded region indicates a prominent structure outside the single image plane. The small arrowheads point to thin tubules that appear to interconnect the more prominent vesicular structures. In the right-most panel the arrowhead and arrow illustrate structures localized at different Z-levels that can be appreciated by rotating the sum of the sections by 90°. Also see QuickTime Demo 1.

Varsha Patki, et al. Mol Biol Cell. 2001 January;12(1):129-141.
11.
Figure 2

Figure 2. From: Insulin Action on GLUT4 Traffic Visualized in Single 3T3-L1 Adipocytes by Using Ultra-fast Microscopy.

Time course of insulin action on GLUT4 accumulation at the plasma membrane. (A) Panels represent a single 250-nm section through the middle of the cell acquired at the times (Min et al., 1999) after insulin addition indicated in the top left corner. A phase image of the cell is shown in the first panel. (B) A rim of 5 pixels around the cell periphery was selected (insets), and the number of pixels of highest intensity (level 153) was calculated using the histogram function in Photoshop 5.0 The numbers at the top left corner represent minutes of insulin treatment. (C) Number of pixels was plotted as a function of time of insulin exposure (see also QuickTime Demo 1).

Varsha Patki, et al. Mol Biol Cell. 2001 January;12(1):129-141.

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