Autophagy is a crucial intracellular pathway that manages the degradation and recycling of long-lived proteins in the cell. The LC3 (or light chain 3) family is composed of three members, LC3A, LC3B and LC3C. Upon autophagy induction, LC3 is cleaved, causing the release of a C-terminal glycine that is required for phospholipid conjugation. This process is vital to the formation of the autophagosome, a double membrane structure that delivers proteins to the lysosome during autophagy. This process has made LC3 a strong indicator of autophagy; therefore, the use of LC3 antibodies in autophagy research is extremely efficient. LC3 and autophagy are important parts of development, differentiation, survival and homeostasis. A link between autophagy and hepatology has been previously established and found to have anywhere from an anti-tumor effect to a pro-survival activity during liver ischemia characterized by nutrient starvation and anoxia.
Denaes et al used an LC3B antibody in their research on the cannabinoid receptor 2 and its protective role against alcoholic liver disease. To begin, this group presented information that the Cannabinoid Receptor 2 (CB2) protects the liver against alcohol-induced disease by regulation of Kupffer cell activation. It was hypothesized that the method of activation of the Kupffer cell reaction was due, in part, to an autophagic response. First, RAW264.7 macrophage cells were exposed to the lysosomal inhibitor chloroquine, and then LC3B expression was measured using an LC3 antibody in western blot. Next, CB2 deficient macrophages were measured for LC3 and SQSTM1/p62 using an LC3B antibody and an SQSTM1/p62 antibody in immunofluorescence. These CB2 deficient macrophages had increased LC3 puncta, pointing to a potential lysosomal defect. Furthermore, an LC3B antibody helped to establish that the CB2 receptor activates autophagy through an HO-1 dependent pathway.
Thapaliya et al used an LC3B antibody to investigate the role that alcohol induced autophagy has in the loss of skeletal muscle mass. For starters, an LC3B antibody was used in western blot on skeletal muscle lysates from alcoholic cirrhotics and control individuals. It was observed that an increase in LC3B lipidation was present in the alcoholic cirrhotics samples. What’s more, western blots using an LC3B antibody of the gastrocnemius muscle showed that ethanol-fed mice had lower ubiquinated proteins compared to their controls. Additionally, an LC3B antibody confirmed that autophagy is induced by the ethanol metabolic acetaldehyde. While these findings do not fully elucidate the role of ethanol-induced autophagy in muscle cells, they do lay the groundwork for the molecular basis.
Novus Biologicals offers LC3B reagents for your research needs including:
PMID: 24492484
PMID: 27346657
PMID: 20810185
