Phorbol ester- and PACAP-induced translocation of PKCγ-GFP fusion
protein in cortical cells. (a) PKCγ-GFP fusion protein
was visualized throughout the cytoplasm in transfected cortical cells.
Note the lack of staining over the eccentric nuclei as indicated by
open arrowheads. This pattern was observed in most unstimulated
cortical cells transfected with PKCγ-GFP alone,
PKCγ-GFP/PAC1short, or PKCγ-GFP/PAC1hop. (b)
Activation of PKCγ by 0.2 μM PMA for 15 min induced the
translocation of PKCγ-GFP from cytosol to the nucleus (indicated by
solid arrowheads). (c) PKCγ-GFP was also visualized
throughout the cytoplasm in PKCγ-GFP/PAC1short-transfected cortical
cells 45 s after 10 nM PACAP treatment. Open arrowhead indicates
absence of prominent nuclear staining. (d) In contrast,
PKCγ translocation to the membrane (arrows) and nucleus (solid
arrowheads) occurred in PKCγ-GFP/PAC1hop-transfected cells 45
s after 10 nM PACAP. (e) Quantification of PKCγ-GFP
nuclear/membrane localization in cells transfected with PKCγ-GFP
alone, PKCγ-GFP/PAC1short, or PKCγ-GFP/PAC1hop after 45 s
PACAP treatment. Data were obtained from three experiments; *,
P < 0.001 vs. GFP-PKC-transfected cells. (Scale
bar, 10 μm.)