Detection of the steroid receptor interacting protein, PAK6, in a neuronal cell line

PAK6 is a Group II p21 activated kinase that unlike traditional signal transduction proteins interacts with multiple binding partners including sex-steroid receptors. PAK6 acts as a nodal checkpoint integrating multiple cellular inputs to promote distinct cellular outcomes, some of which are associa...

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Main Authors: Sharon Ramos Goyette, Eric Schott, Astopheline Uwimana, David W. Nelson, Jacob Boganski
Format: Article
Language:English
Published: Elsevier 2019-03-01
Series:Heliyon
Subjects:
Online Access:http://www.sciencedirect.com/science/article/pii/S2405844018346188
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spelling doaj-5c3842a3683a4cffb8462b7656631aa52020-11-25T02:02:24ZengElsevierHeliyon2405-84402019-03-0153e01294Detection of the steroid receptor interacting protein, PAK6, in a neuronal cell lineSharon Ramos Goyette0Eric Schott1Astopheline Uwimana2David W. Nelson3Jacob Boganski4Corresponding author.; Shields Science Center, Stonehill College, Easton MA, 02357, USAShields Science Center, Stonehill College, Easton MA, 02357, USAShields Science Center, Stonehill College, Easton MA, 02357, USAShields Science Center, Stonehill College, Easton MA, 02357, USAShields Science Center, Stonehill College, Easton MA, 02357, USAPAK6 is a Group II p21 activated kinase that unlike traditional signal transduction proteins interacts with multiple binding partners including sex-steroid receptors. PAK6 acts as a nodal checkpoint integrating multiple cellular inputs to promote distinct cellular outcomes, some of which are associated with cytoskeletal remodeling. Despite the possibility that PAK6 may couple sex-specific neuronal function and therefore serve as a valuable research, diagnostic and therapeutic target, there is currently no standardized protocol for assessing PAK6 activity in a neuronal cell line. Here, we present a protocol for assessing PAK6 levels in a commonly used neuronal cell line, PC-12. In comparison with other methodology, this approach (1) does not require ex-planted tissue to identify PAK6 in neurons and (2) unlike other protocols which require steroid depleted media for detection of PAK6 in non-neuronal cell lines, such as prostate cancer cell lines, we were easily able to detect PAK6 in PC-12 cells grown in complete, steroid-containing media. Thus the present protocol allows for the efficient detection of native PAK6 in PC-12 cells to expedite targeted basic research of the emerging importance of PAK6 function in the brain as well as to accelerate the identification and isolation of potential therapeutic targets not only in cancerous but brain disease states as well.http://www.sciencedirect.com/science/article/pii/S2405844018346188Cancer researchCell biologyNeuroscience
collection DOAJ
language English
format Article
sources DOAJ
author Sharon Ramos Goyette
Eric Schott
Astopheline Uwimana
David W. Nelson
Jacob Boganski
spellingShingle Sharon Ramos Goyette
Eric Schott
Astopheline Uwimana
David W. Nelson
Jacob Boganski
Detection of the steroid receptor interacting protein, PAK6, in a neuronal cell line
Heliyon
Cancer research
Cell biology
Neuroscience
author_facet Sharon Ramos Goyette
Eric Schott
Astopheline Uwimana
David W. Nelson
Jacob Boganski
author_sort Sharon Ramos Goyette
title Detection of the steroid receptor interacting protein, PAK6, in a neuronal cell line
title_short Detection of the steroid receptor interacting protein, PAK6, in a neuronal cell line
title_full Detection of the steroid receptor interacting protein, PAK6, in a neuronal cell line
title_fullStr Detection of the steroid receptor interacting protein, PAK6, in a neuronal cell line
title_full_unstemmed Detection of the steroid receptor interacting protein, PAK6, in a neuronal cell line
title_sort detection of the steroid receptor interacting protein, pak6, in a neuronal cell line
publisher Elsevier
series Heliyon
issn 2405-8440
publishDate 2019-03-01
description PAK6 is a Group II p21 activated kinase that unlike traditional signal transduction proteins interacts with multiple binding partners including sex-steroid receptors. PAK6 acts as a nodal checkpoint integrating multiple cellular inputs to promote distinct cellular outcomes, some of which are associated with cytoskeletal remodeling. Despite the possibility that PAK6 may couple sex-specific neuronal function and therefore serve as a valuable research, diagnostic and therapeutic target, there is currently no standardized protocol for assessing PAK6 activity in a neuronal cell line. Here, we present a protocol for assessing PAK6 levels in a commonly used neuronal cell line, PC-12. In comparison with other methodology, this approach (1) does not require ex-planted tissue to identify PAK6 in neurons and (2) unlike other protocols which require steroid depleted media for detection of PAK6 in non-neuronal cell lines, such as prostate cancer cell lines, we were easily able to detect PAK6 in PC-12 cells grown in complete, steroid-containing media. Thus the present protocol allows for the efficient detection of native PAK6 in PC-12 cells to expedite targeted basic research of the emerging importance of PAK6 function in the brain as well as to accelerate the identification and isolation of potential therapeutic targets not only in cancerous but brain disease states as well.
topic Cancer research
Cell biology
Neuroscience
url http://www.sciencedirect.com/science/article/pii/S2405844018346188
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