Trypsin-induced depletion of plasma membrane PIP2 and
formation of plasma membrane DAG and cytoplasmic IP3.
A, confocal fluorescence images of the GFP-PH probe for
PIP2/IP3 before (a) and after (b) 1
μm trypsin treatment and of the YFP-C1 probe for DAG before
(c) and after (d) trypsin treatment. B, time
courses of cytoplasmic fluorescence, normalized to the initial fluorescence
(F/Fo). C, average cytoplasmic fluorescence ratio
(F/Fo), with F being the maximal fluorescence
after trypsin treatment (only responsive cells were analyzed: 6/6 cells for
GFP-PH and 9/12 cells for YFP-C1). D, fluorescence images of the
GFP-PH probe before (a) and after (b) 100 μm
PAR-2 AP stimulation and of the YFP-C1 probe before (c) and after
(d) PAR-2 AP. E, time course of cytoplasmic ratio.
F, average cytoplasmic fluorescence ratio after PAR-2 AP treatment;
the majority of tested cells showed reciprocal translocation of GFP-PH (7/8
cells) and YFP-C1 (3/4).