Figure 4.
cAMP-dependent translocation of sGC to the membrane. Starved sGC-GFPoe cells were settled in a perfusion chamber, stimulated with 10-6 M cAMP at t = 0 s, and a sequence of images was recorded with an exposure time of 2 s per frame. (A) A cell just before cAMP stimulation and at 10 s after stimulation. (B) The fluorescence intensity of sGC-GFP in the cytosol after cAMP stimulation is presented relative to the cytosolic fluorescence intensity before cAMP stimulation. (C) Translocation of sGC activity. Cells in suspension were stimulated at t = 0 s with 0.1 μM cAMP at 22°C and lysed at 0°C at the indicated time points. The lysate was centrifuged and Mn2+-dependent sGC activity was determined in the pellets (filled bars) and supernatants (open bars on top of filled bars). The activity in the pellet at t = 10 s is significantly higher compared with t = 0 s (* Student's t test at p < 0.05); at t = 60 s the difference is not significant (ns). The data shown are the means and SE of the means of two experiments each performed with lysis in triplicate.