Figure - PMC

Skip to main content

An official website of the United States government

Here's how you know

Here's how you know

Official websites use .gov
A .gov website belongs to an official government organization in the United States.

Secure .gov websites use HTTPS
A lock ( ) or https:// means you've safely connected to the .gov website. Share sensitive information only on official, secure websites.

View full-text article in PMC

. 1997 Apr 21;137(2):481–492. doi: 10.1083/jcb.137.2.481

Search in PMC
Search in PubMed
View in NLM Catalog
Add to search

This article is distributed under the terms of an Attribution–Noncommercial–Share Alike–No Mirror Sites license for the first six months after the publication date (see http://www.rupress.org/terms). After six months it is available under a Creative Commons License (Attribution–Noncommercial–Share Alike 4.0 Unported license, as described at http://creativecommons.org/licenses/by-nc-sa/4.0/).

PMC Copyright notice

MLCK activity is required for MAP kinase-induced phosphorylation of MLC and cell migration. (A, upper panel) COS cells were transfected with empty vector or mutationally active MEK+ along with an HA-tagged ERK2 reporter construct. Cells were allowed to migrate on a collagen substrate for 3 h in the presence or absence of the MLCK inhibitor KT5926 (10 μM) as described in Materials and Methods. Cell migration was enumerated as described above. (Lower panel) Lysates prepared from these COS cells metabolically labeled with ³²P were examined by SDS-PAGE and autoradiography. The position of HA-tagged ERK and MLC are denoted by arrows. (B, upper panel) COS cells were transfected with mutationally active MEK+ and/or MLCK(Mut) and allowed to migrate on a collagen substrate as described in Materials and Methods. Cell migration was quantified by counting the number of migrant cells per high powered microscopic field as described above. (Lower panel) Lysates prepared from ³²P metabolically labeled COS cells transfected as above were examined for phosphorylation of HA-ERK and MLC. The result shown is a representative experiment from at least three independent experiments.

MLCK activity is required for MAP kinase-induced phosphorylation of MLC and cell migration. (A, upper panel) COS cells were transfected with empty vector or mutationally active MEK+ along with an HA-tagged ERK2 reporter construct. Cells were allowed to migrate on a collagen substrate for 3 h in the presence or absence of the MLCK inhibitor KT5926 (10 μM) as described in Materials and Methods. Cell migration was enumerated as described above. (Lower panel) Lysates prepared from these COS cells metabolically labeled with ³²P were examined by SDS-PAGE and autoradiography. The position of HA-tagged ERK and MLC are denoted by arrows. (B, upper panel) COS cells were transfected with mutationally active MEK+ and/or MLCK(Mut) and allowed to migrate on a collagen substrate as described in Materials and Methods. Cell migration was quantified by counting the number of migrant cells per high powered microscopic field as described above. (Lower panel) Lysates prepared from ³²P metabolically labeled COS cells transfected as above were examined for phosphorylation of HA-ERK and MLC. The result shown is a representative experiment from at least three independent experiments.