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Fig. 6 | Cell & Bioscience

Fig. 6

From: β-amyloid protein induces mitophagy-dependent ferroptosis through the CD36/PINK/PARKIN pathway leading to blood–brain barrier destruction in Alzheimer’s disease

Fig. 6

A low dose of Aβ1-40 causes cellular mitophagy-dependent lipid peroxidation and induces ferroptosis through CD36/PINK1/Parkin pathway. A The GSH-PX kit was used to detect the changes in pericyte glutathione peroxidase stimulated by Aβ1-40. B The iron kit detects changes in iron ions. C A fluorescence microplate reader (Mito-FerroGreen) was used to detect the change in Fe2 + . D Fluorescence microplate reader analysis of BODIPY581/591 detects the effect of lipid active oxygen. E Western blot analysis was used to detect changes in the expression of the ferroptosis-related proteins GPx4, xCT, ferritin, and NOX1 after stimulation of pericytes with 1 µM Aβ1-40 or 10 µM erastin for 6 h. F–I Semiquantitative analysis of GPx4, xCT, ferritin, and NOX1 protein levels. The data are presented as the means ± SD (n = 3 per group). J Western blotting was used to detect the changes in the expression of the ferroptosis-related proteins GPx4, xCT, ferritin, and NOX1 after pericytes were stimulated with 1 µM Aβ1-40 with or without FER-1 for 6 h. K–N Semiquantitative analysis of GPx4, xCT, ferritin, and NOX1 protein levels. The data are presented as the means ± SD (n = 3 per group). T Erastin (10 µM) was used as a positive control; typical transmission electron micrograph after 6 h with or without FER-1 (10 µM). At least 6 cells were examined under each treatment condition. O Erastin (10 µM) was used as a positive control; typical transmission electron micrograph after 6 h with or without FER-1 (10 µM). At least 6 cells were examined under each treatment condition. PS Semiquantitative analysis of GPx4, xCT, ferritin, and NOX1 protein levels. The data are presented as the means ± SD (n = 3 per group). U Western blotting was used to detect the expression levels of the mitochondrial pathway proteins S65, BNIP3/NIX, and PINK1/Parkin after 1 µM Aβ1-40 treatment of pericytes for 6 h. VZ Semiquantitative analysis of S65, BNIP3/NIX, and PINK1/Parkin protein levels. The data are presented as the means ± SD (n = 6 per group). a Western blot detection of the effect of 1 µM Aβ1-40 on the expression of CD36, PINK1/Parkin, HSP60, and Tim23 in pericytes 6 h after si-CD36 transfection. bd Semiquantitative analysis of CD36, PINK1/Parkin, HSP60, and Tim23 protein levels. The data are presented as the means ± SD (n = 3 per group). (*p < 0.05, **p < 0.01, ***p < 0.001 compared with the CTR group, &p < 0.05, &&p < 0.01 compared with the 1 µM Aβ1-40 group, n = 3 or 6)

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