![Fig. 5. Forskolin-treatment of Chinese hamster ovary cells expressing wild-type GpIb/β/IX enhanced the interaction between 14-3-3ζ and the GpIb complex. / CHO β/IX cells expressing vector, wild-type GpIbα or GpIbα deleted from residues 570-590 were treated for 1 hour with 100 μmol/L forskolin, which stimulated cyclic adenosine monophosphate-dependent protein kinase. (A) Forskolin caused increased phosphorylation of recombinant GpIbβ in CHO cells radiolabeled with 32P] orthophosphate. (B) Forskolin increased the amount of recombinant GpIbβ that bound to the GST-14-3-3ζ fusion protein only in cells expressing wild-type GpIbα. (C) Forskolin increased the quantity of 14-3-3ζ that coimmunoprecipitated with recombinant GpIbα from CHO cells transduced with wild-type GpIbα, GpIbβ, and GpIX. (D) Quantitative analysis of the effect of forskolin-induced PKA activation on the amount of 14-3-3ζ that coimmunoprecipitated with GpIbα and the amount of GpIbβ and GpIbα that bound to the GST-14-3-3ζ fusion protein, in CHO cells stably expressing wild-type GpIbα, GpIbβ, and GpIX. The amount of protein is reported as arbitrary densitometry units. (n = 3; *P < .001, Student ttest).](https://ash.silverchair-cdn.com/ash/content_public/journal/blood/95/2/10.1182_blood.v95.2.551/6/bloo00227005dx.jpeg?Expires=1723372313&Signature=SBSCk2IQQp3uhS7QN2eBOOuyWJtErArwIuy6MLGBFXFZLqIebugtWSfNPNX6~FWo9n0bGoGMDzF0NSWdFAPlKb3CXvMNIwg0tZ4W3drOYaIES2qdhI9SIFg0LqwpJn0gkKbrSJn3Hkx8xqx-3KL8QUW7EafXd2aCQsUzZ-2-ZMqDT98NkQMU5Zz3KoI1uNy8ElUeedLcmMm2UUkP-Ty8QF23JxBLC~SHHsGBKbDfs85tf4NJ7O5LNEHxPfPDhD6dZEz3PYFsy1EYdf1LBh4WtaWQKb-9WXQQpbf692e6vMH~S6rBYnrvz3xW9tKL0tFPPCaZwwfCsDU1nxEUdX~eRA__&Key-Pair-Id=APKAIE5G5CRDK6RD3PGA)
Forskolin-treatment of Chinese hamster ovary cells expressing wild-type GpIb/β/IX enhanced the interaction between 14-3-3ζ and the GpIb complex.
CHO β/IX cells expressing vector, wild-type GpIbα or GpIbα deleted from residues 570-590 were treated for 1 hour with 100 μmol/L forskolin, which stimulated cyclic adenosine monophosphate-dependent protein kinase. (A) Forskolin caused increased phosphorylation of recombinant GpIbβ in CHO cells radiolabeled with 32P] orthophosphate. (B) Forskolin increased the amount of recombinant GpIbβ that bound to the GST-14-3-3ζ fusion protein only in cells expressing wild-type GpIbα. (C) Forskolin increased the quantity of 14-3-3ζ that coimmunoprecipitated with recombinant GpIbα from CHO cells transduced with wild-type GpIbα, GpIbβ, and GpIX. (D) Quantitative analysis of the effect of forskolin-induced PKA activation on the amount of 14-3-3ζ that coimmunoprecipitated with GpIbα and the amount of GpIbβ and GpIbα that bound to the GST-14-3-3ζ fusion protein, in CHO cells stably expressing wild-type GpIbα, GpIbβ, and GpIX. The amount of protein is reported as arbitrary densitometry units. (n = 3; *P < .001, Student ttest).
