Abstract
The LC3B protein plays a critical role in autophagy. Normally, this protein resides in the cytosol, but following cleavage and lipidation with phosphatidylethanolamine, LC3B associates with the phagophore. This localization can be used as a general marker for autophagic membranes. To visualize the LC3B, we used the LC3B Antibody Kit for Autophagy (Invitrogen). As this kit was designed to work with cell coming from cell culture it was not possible to perform it on compact tissues such as nerve cord or ovaries. We thus adapted LC3B labeling initial protocol to be able to label autophagic membranes of adipocytes from terrestrial isopods which form a loose tissue that can be more easily penetrated by antibodies. The following protocol permits a visualization of the autophagic vesicles directly in terrestrial isopod cells freshly sampled in animals.
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