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  • In Vitro Explant Assays and Cultures to Study PCP Signaling in Axon Guidance.

In Vitro Explant Assays and Cultures to Study PCP Signaling in Axon Guidance.

Methods in molecular biology (Clifton, N.J.) (2022-02-12)
Pau Marfull-Oromí, Kathryn M Miller, Yimin Zou
ABSTRACT

In vitro studies have provided valuable insights to the function and mechanisms in axon guidance. In this chapter, we will introduce the rodent "open-book" assay, pre- or postcrossing explant culture and the dissociated neuron culture. They have been used to discover mechanism which we have gone on to validate or will confirm using in vivo genetic approaches.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Forskolin, For use in molecular biology applications
Sigma-Aldrich
Gö 6976, Gö 6976, CAS 136194-77-9, is a cell-permeable, reversible, and ATP-competitive inhibitor of PKC (IC₅₀ = 7.9 nM for rat brain). Exhibits selectively for PKCα (IC₅₀ = 2.3 nM) and βI (IC₅₀ = 6.2 nM).
Roche
Collagen, from rat tail tendon
Sigma-Aldrich
Lithium chloride, BioXtra, ≥99.0% (titration)
Sigma-Aldrich
SB 216763, >98% (HPLC)
Sigma-Aldrich
JNK Inhibitor I, (L)-Form, Cell-Permeable, The JNK Inhibitor I, (L)-Form, Cell-Permeable controls the biological activity of JNK. This small molecule/inhibitor is primarily used for Phosphorylation & Dephosphorylation applications.