Abstract：Abstract: In order to facilitate the high throughput screening of autophagy related drugs on islet β cells, the RFP-GFP-LC3 plasmid was transfected into RIN-m5f cells by lentivirus transfection, and the infected cells were screened with G418. Laser confocal imaging verified that a cell line expressing RFP-GFP-LC3 was obtained. After different treatments of RIN-m5f, the results showed that the fluorescence point ratio of GFP/RFP in the starved group was lower than that in the normal group, and that expression of P62 protein and the level of S6K phosphorylation were decreased. After 10 μmol/L chloroquine was added to partially inhibit autophagy, the fluorescence point ratio of GFP/RFP increased. These results indicated that the rat islet β cell strain stably expressing RFP-GFP-LC3 was successfully constructed. The cells could exhibit the whole process of autophagy in a simple and accurate manner, which lays a foundation for the study of mechanisms of autophagy related to diabetic medicines.
引用本文:
周佳丽, 吴艳阳, 罗玉霜, 袁玉菊, 刘明俊, 刘东波. 稳定表达RFP-GFP-LC3的大鼠胰岛β细胞株的构建[J]. 生命科学研究, 2019, 23(2): 87-91. ZHOU Jia-li, WU Yan-yang, LUO Yu-shuang, YUAN Yu-ju, LIU Ming-jun, LIU Dong-bo. Establishment of RIN-m5f Cell Strain Stably Expressing RFP-GFP-LC3. Life Science Research, 2019, 23(2): 87-91.
