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. Author manuscript; available in PMC: 2009 Feb 12.
Published in final edited form as: Curr Biol. 2008 Jan 31;18(3):177–182. doi: 10.1016/j.cub.2007.12.050

Figure 3. Analysis of GFP-STIM1 dynamics in control cells and after Ca2+ store depletion.

Figure 3

A. FRAP analysis of GFP-STIM1 behavior. Each panel, with the exception of the panel marked “FRAP” (which shows a single frame), represents superimposition of 5 successive frames with a 1s interval. Note that recovery of diffuse ER signal in the bleached area precedes the appearance of GFP-STIM1 comets (indicated by red arrows).

B. The average GFP-STIM1 intensity ratio of two regions inside and outside of the photobleached area in HeLa cells (measured as described in [14]). Left: control cells, n = 20; cells after addition of 2 μM thapsigargin (TG), n = 13 cells; middle: control cells, n = 7; 2 μm thapsigargin n = 12 cells; right: GFP-STIM, n = 20 cells; EB3-mRFP, n = 7 cells.

C. Representative frames of simultaneous two-color video of an MRC5-SV cell expressing GFP-STIM1 and EB3-RFP before and 120s after the addition of 2 μM thapsigargin in normal culture medium. Kymographs illustrating the changes of fluorescent intensity over time in the indicated boxed areas are shown on the right. In kymographs motile comets appear as slopes and immobile structures as vertical lines. Bars, 5 μm.