Figure 5. ER sheets spread into the cell periphery when Rab18 or its GEF complex is depleted. (a) COS7 cells were depleted of Rab3GAP subunits (GAP1 and GAP2), Rab18, or Rab10 for 72 h. Two different siRNA duplexes to Rab3GAP1 (GAP1.1 and GAP1.2) were used. The cells were fixed and then stained with antibodies to CLIMP-63 and reticulon 4 (Rtn4). DNA was stained with DAPI (blue in the merged panel). Dotted yellow lines mark the cell boundaries. The bar is marked in the figure. (b) The area of CLIMP-63 as a function of total cell area was measured using ImageJ (National Institutes of Health) for 50–70 cells per experiment, for three independent experiments (blue bars). Additionally, the percentage of cells showing spread CLIMP-63 was also counted (green bars). In both cases the mean values are plotted in the bar graph, with error bars indicating the standard deviation of the mean.
Figure 5.

ER sheets spread into the cell periphery when Rab18 or its GEF complex is depleted. (a) COS7 cells were depleted of Rab3GAP subunits (GAP1 and GAP2), Rab18, or Rab10 for 72 h. Two different siRNA duplexes to Rab3GAP1 (GAP1.1 and GAP1.2) were used. The cells were fixed and then stained with antibodies to CLIMP-63 and reticulon 4 (Rtn4). DNA was stained with DAPI (blue in the merged panel). Dotted yellow lines mark the cell boundaries. The bar is marked in the figure. (b) The area of CLIMP-63 as a function of total cell area was measured using ImageJ (National Institutes of Health) for 50–70 cells per experiment, for three independent experiments (blue bars). Additionally, the percentage of cells showing spread CLIMP-63 was also counted (green bars). In both cases the mean values are plotted in the bar graph, with error bars indicating the standard deviation of the mean.

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