%0 Journal Article
%T Suppression of astrocytic autophagy by ¦ÁB-crystallin contributes to ¦Á-synuclein inclusion formation
%A Fei Ding
%A Jia-wei Zhou
%A Pei-zhou Liang
%A Shen-zhao Lu
%A Shu Yin
%A Shu-zhen Zhang
%A Xiao-min Wang
%A Xiao-song Gu
%A Yan-qing Yin
%A Yong-shun Guo
%J Archive of "Translational Neurodegeneration".
%D 2019
%R 10.1186/s40035-018-0143-7
%X CRYAB regulates autophagy process in vitro. a Knockdown of CRYAB enhances the conversion of LC3-I to LC3-II in U251 cells. U251 cells were transfected with CRYAB siRNA (siNC: the siRNA of negative control; siCRYAB: the siRNA of CRYAB) and the cellular LC3 levels were assessed by Western blotting 48£¿h after transfection. Data are presented as mean£¿¡À£¿SEM, n£¿=£¿6. Unpaired t-test. **P£¿<£¿0.01. b Overexpression of CRYAB inhibits LC3-II formation in U251 cells. The cellular LC3 levels were assessed by western blotting 48£¿h after transfection. Data are presented as the mean£¿¡À£¿SEM, n£¿=£¿3. Unpaired t-test. **P£¿<£¿0.01. c CRYAB overexpression in primary astrocytes inhibits LC3-II formation. Primary cultured astrocytes were transfected with lentivirus vector encoding CRYAB. The cellular LC3 levels were assessed by Western blotting 48£¿h after transfection. Data are presented as mean£¿¡À£¿SEM, n£¿=£¿3. Paired t-test. *P£¿<£¿0.05. d U251 cells were transfected with siCRYAB in the presence or absence of H2O2 (1£¿mM, 6£¿h). The cellular p62 levels were assessed 48£¿h after transfection using western blotting. Data are presented as mean£¿¡À£¿SEM, n£¿=£¿3. *P£¿<£¿0.05. e Knockdown of CRYAB leads to a marked increase in the number of autophagic vacuoles. Autophagic vacuoles in U251 cells transfected with siNC or siCRYAB were examined using electron microscopy. The arrows indicate autophagic vacuoles. Scale bars: 0.5£¿¦Ìm. The graph shows a statistical analysis of cytoplasmic occupancy of autophagic vacuoles in the cells. Data are presented as mean£¿¡À£¿SEM. Twenty cells in each group were examined. Unpaired t-test. ***P£¿<£¿0.00
%K ¦ÁB-crystallin
%K Astrocytes
%K Parkinson¡¯s disease
%K Autophagy
%U https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6337871/