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Taking a roller coaster ride with autophagy markers p62 and LC3
Author(s) -
Koskela A.,
Reinisalo M.,
Kaarniranta K.
Publication year - 2016
Publication title -
acta ophthalmologica
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.534
H-Index - 87
eISSN - 1755-3768
pISSN - 1755-375X
DOI - 10.1111/j.1755-3768.2016.0539
Subject(s) - autophagy , microbiology and biotechnology , western blot , homeostasis , biology , organelle , biochemistry , apoptosis , gene
Purpose Lysosomal autophagy is crucial for the removal of dysfunctional proteins, aggregated cellular material and organelles. Post‐mitotic cells, such as retinal pigment epithelial cells, are particularly susceptible to age‐related aggregation‐prone proteins. Excessive or defective activity of the autophagy leads to detrimental effects on RPE cell homeostasis. Therefore, autophagy must be controlled not only with positive but also with negative signals to retain homeostasis. In this work, autophagy markers LC3 (microtubule‐associated protein 1A/1B‐light chain 3) and p62 (SQSTM1) were monitored in ARPE‐19 cell line. Methods ARPE‐19 cells were treated with autophagy inducers, proteasome inhibitor MG‐132 and Resvega (contains resveratrol and omega‐3 fatty acids), in normal and serum‐starved growth conditions up to 48 h. Protein samples from different time points were collected and the protein levels of LC3‐II, LC3‐I and p62 were analyzed with Western blot. Results The levels of LC3‐II, LC3‐I and p62 fluctuated during the monitored time period in both growth conditions. The magnitude of the fluctuation was highest in the serum‐starved samples at early time points and became weaker at the later time points. The expression levels of autophagy markers were constant throughout the experiment in normal growth conditions. Conclusions This data showed that autophagy regulation is a dynamic process that coincides with changed and time dependent expression levels of LC3 and p62 in response to MG‐132, Resvega and starvation. This should be noticed when autophagy data is analyzed.