The autophagy related gene 8 (Atg8) family proteins play a central role at various steps of autophagy, including autophagosome initiation, elongation, maturation and lysosome fusion, as well as cargo targeting. However, details regarding the distinct roles of individual Atg8 members during autophagy is not much known.
The objective of this project is to determine the roles of each Atg8 family protein and understand their intracellular signaling and molecular effects during autophagy induced by 2-deoxy-D-glucose (2-DG) treatment in adult hippocampal neural stem (HCN) cells. Especially, we will be focusing on three proteins: microtubule-associated protein 1 light chain 3 beta (LC3B), GABA type A receptor associated protein like 1 (GABARAPL1), and 2 (GABARAPL2). HCN cells with stable knockdown of individual Atg8 family member proteins will be treated with 2-DG to induce autophagy and downstream signaling pathways will be examined.
Through these experiments, we will elucidate the effects of Atg8 family proteins and further understand the relevance of Atg8 proteins during autophagy in HCN cells following 2-DG treatment. ⓒ 2017 DGIST
Table Of Contents
1. Introduction 5--
1.1 General introduction to autophagy 5--
1.2 Adult hippocampal neural stem cells 5--
1.3 ATG8 family members 8--
1.4 2-deoxy-D-glucose 12--
2. Materials & Methods 14--
2.1 Cell culture & Atg8 member knockdown cell 14--
2.2 Antibody and Chemicals 14--
2.3 Cell Death Assay 15--
2.4 2DG drug preparation and treatment 15--
2.5 Western Blot 16--
3. Results 17--
3.1 Effects of Atg8 knockdowns on HCN cell survival 17--
3.2 Responses of Atg8 family members upon 2DH treatment 18--
3.3 Effects of Atg8 knockdown in HCN cells upon 2DG treatment 20--