Fig. 5
Autophagic agonists and inhibitors verified that 1 µM Aβ1-40 treatment of pericytes led to increased mitochondrial autophagy and increased the generation of ROS. A Western blot detection of the mitochondrial proteins Tim23 and HSP60 and the autophagosome protein LC3II/LC3I at the protein level after 5 µM CCCP treatment as a positive control. B–D Semiquantitative analysis of Tim23, HSP60, and LC3II/LC3I protein levels. The data are presented as the means ± SD (n = 3 per group). E Western blots to detect the expression changes in the mitochondrial proteins Tim23 and HSP60 at the protein level with or without 20 µM CQ. F, G Semiquantitative analysis of Tim23 and HSP60 protein levels. The data are presented as the means ± SD (n = 3 per group). (H) Western blot to detect the expression changes of mitochondrial proteins Tim23 and HSP60 at the protein level with or without Mdivi-1 20 µM. I, J Semiquantitative analysis of Tim23 and HSP60 protein levels. The data are presented as the means ± SD (n = 3 per group). (**p < 0.01 compared with the CTR group, &&p < 0.01 compared with the 1 µM Aβ1-40 group, n = 3). K Use of 5 µM CCCP as a positive control and typical transmission electron micrographs after 6 h with or without 20 µM Mdivi-1. At least 6 cells were examined under each treatment condition. L Western blotting was used to detect changes in the expression of apoptosis-related BCL2/BAX and P17/P35 CASP3 after Aβ1-40 (100 nM and 1 µM) stimulation of pericytes for 6 h. M, N Semiquantitative analysis of BCL2/BAX and P17/P35 CASP3 protein levels. The data are presented as the means ± SD (n = 3 per group). O Flow cytometry to detect the effect of Aβ1-40 on pericyte apoptosis. P Fluorescence microscopy observation and detection of the fluorescent probe DHE indicate that Aβ1-40 stimulation changes ROS expression in pericytes. Q Fluorescence microscopy observation and detection of the fluorescent probe for ROS, which is used to show the changes in ROS expression in pericytes caused by Aβ1-40 stimulation. R The ATP kit detects the changes in pericyte energy produced by Aβ1-40 stimulation. S SOD kit detection of the effect of Aβ1-40 stimulation on the antioxidant capacity of pericytes (*p < 0.05, **p < 0.01 and CTR group, n = 3)