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Integrins and Signal Transduction
We will inform you when it is available. Crk p Nck1 NCK2. Actin Dynamics Signaling Pathway Background. Signaling Pathways. Product type.
Research area. About us. Lee, J. Rubin, S.
Purpose: : To elucidate molecular mechanisms of myocilin Myoc action. Methods: : Conditioned media was prepared from HEK cells that were transiently transfected with cDNAs encoding wild-type or mutated human Myoc and its N-terminal and C-terminal fragments. Flag-tagged Myoc was purified using Flag-agarose.
Actin cytoskeleton was visualized by staining with Rhodamin-phalloidin. Flag- and alkaline phosphatase-tagged Myoc were used to study its interaction with frizzed receptors and secreted frizzled-related proteins sFRPs. Results: : Addition of conditioned media from Myoc expressing cells or purified Myoc protein to NIH3T3 or human trabecular meshwork cells induced changes in their actin cytoskeleton similar to those observed after addition of Wnt3a conditioned media. Conditioned media from cells expressing non-secreted mutated Myoc did not induce changes in the actin cytoskeleton.
Actin Dynamics Signaling Pathway
Pre-incubation of conditioned media from Myoc expressing cells but not from Wnt-expressing cells with monoclonal antibodies against Myoc blocked changes in actin cytoskeleton. Changes in the actin cytoskeleton were also blocked by addition of Wnt signaling antagonist, sFRP1, to conditioned media.
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Direct interaction of Myoc with sFRP1 was confirmed in co-immunoprecipitation experiments. The olfactomedin domain of Myoc was critical for this interaction.
Alkaline phosphatase-tagged human Myoc interacted with several frizzled receptors and sFRPs with frizzled-7 and sFRP3 giving the highest affinity of binding. Conclusions: : Myoc may be involved in the regulation of actin cytoskeleton in the trabecular meshwork through components of Wnt signaling pathway.
Our data suggest that Myoc may work as a modulator of Wnt signaling. Purchase this article with an account. Kwon ; H. Lee ; J.
Related Signaling and the Cytoskeleton
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