Effects of InsP3 buffering on AngII-induced Tubby-domain and PLCδ1PH translocation responses. HEK293-AT1 cells were transfected with the indicated GFP reporter constructs alone or together with p130PH-mRFP. After 24 hours, cells were examined in a Zeiss Live 5 DuoScan confocal microscope at room temperature. Cells were stimulated with 100 nM AngII at 60 second followed by 10 μM ionomycin at 360 sec. Data were collected at 1 fps rate and the cytoplasmic fluorescence intensity was analyzed in selected regions of interest outside the nucleus. Values were normalized for minimum and maximum fluorescence values for each run and these values were averaged. Means ± S.E.M. are shown obtained from: n = 10 (Tubby domain alone), n = 12 (PLCδ1PH alone), n = 21 (Tubby domain + p130PH-mRFP) and n = 20 (PLCδ1PH + p130PH-mRFP) cells. The traces were truncated at 420 sec to better illustrate the AngII-induced changes.
Szentpetery et al. BMC Cell Biology 2009 10:67 doi:10.1186/1471-2121-10-67