Figure 4. ROCK1 generates actomyosin... | Rockefeller University Press

Figure 4.

$Figure 4. ROCK1 generates actomyosin filament bundles that form the cell rear. (A) Confocal images of CHO.K1 cells cotransfected with either ROCK isoform–specific shRNAs or pSUPER control empty vector and Myosin IIB–GFP (MIIB-GFP). The cell edge is outlined in magenta to indicate morphology. (B) Quantification of the ratio between integrated density for MIIB-GFP intensity at the rear (180° ± 90°) versus the front of the cell (0° ± 90°) and the ratio between the two sides of the cell (90° ± 90° and 270° ± 90°), where the integrated density = mean gray value × area. The numerator is defined as the side with increased MIIB-GFP intensity; n = 28 control cells, 21 ROCK1 shRNA cells, and 37 ROCK2 shRNA cells. (C) FRAP recovery curves for photobleached MIIB-GFP in ROCK shRNA and control cells. (D) MIIB mobile fraction calculated from FRAP recovery curves as the percentage of recovered MIIB fluorescence intensity; n = 23 control cells, 13 ROCK1 shRNA cells, and 9 ROCK2 shRNA cells for FRAP data in C and D. Error bars indicate SEM. Ctrl, control.$

ROCK1 generates actomyosin filament bundles that form the cell rear. (A) Confocal images of CHO.K1 cells cotransfected with either ROCK isoform–specific shRNAs or pSUPER control empty vector and Myosin IIB–GFP (MIIB-GFP). The cell edge is outlined in magenta to indicate morphology. (B) Quantification of the ratio between integrated density for MIIB-GFP intensity at the rear (180° ± 90°) versus the front of the cell (0° ± 90°) and the ratio between the two sides of the cell (90° ± 90° and 270° ± 90°), where the integrated density = mean gray value × area. The numerator is defined as the side with increased MIIB-GFP intensity; n = 28 control cells, 21 ROCK1 shRNA cells, and 37 ROCK2 shRNA cells. (C) FRAP recovery curves for photobleached MIIB-GFP in ROCK shRNA and control cells. (D) MIIB mobile fraction calculated from FRAP recovery curves as the percentage of recovered MIIB fluorescence intensity; n = 23 control cells, 13 ROCK1 shRNA cells, and 9 ROCK2 shRNA cells for FRAP data in C and D. Error bars indicate SEM. Ctrl, control.