Examining pi3K-Signaling-Dependent Regulation of Lens Organelle Free Zone Formation via Immunolocalization and Immunoblotting in Chick Embryos

Authors

Rifah Gheyas, Thomas Jefferson UniversityFollow
A. Sue Menko, Thomas Jefferson UniversityFollow

Document Type

Article

Publication Date

9-13-2023

Comments

This article is the author's final published version in STAR Protocols, Volume 4, Issue 4, 2023, Article number 102569.

The published version is available at https://doi.org/10.1016/j.xpro.2023.102569.

Copyright © 2023 The Author(s). This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).

Abstract

The elimination of lens organelles during development, required for mature lens function, is an autophagy-dependent mechanism induced through suppression of PI3K signaling. Here, we present a protocol for investigating the signaling pathways responsible for induction of the formation of this lens organelle free zone. We describe steps for preparation of lens organ culture and use of signaling pathway inhibitors. We then detail procedures for analyzing their impact using both confocal microscopy imaging of immunolabeled lens cryosections and immunoblot approaches. For complete details on the use and execution of this protocol, please refer to Gheyas et al. (2022).

Recommended Citation

Gheyas, Rifah and Menko, A. Sue, "Examining pi3K-Signaling-Dependent Regulation of Lens Organelle Free Zone Formation via Immunolocalization and Immunoblotting in Chick Embryos" (2023). Computational Medicine Center Faculty Papers. Paper 48.
https://jdc.jefferson.edu/tjucompmedctrfp/48

Creative Commons License

This work is licensed under a Creative Commons Attribution 4.0 License.

PubMed ID

37713308

Language

English