Extended Data Fig. 1: MFE-2 defect was related to microglial activation in AD mice.
a. The specificity of MFE-2 antibody used in this study was verified by immunofluorescences of MFE-2 in cultured primary microglia from Flox and ΔMFE-2 mice brain. Experiments were repeated three times. b. Representative RNAscope result of MFE-2 mRNA in microglia of 8-month-old untreated and 5xFAD mouse models (n = 3 per each group, male). Hsd17b4 in red, microglia in green and nuclei in blue. Scale bar, 25 μm. c. Schematic depicting brain microglia isolation by FACS experimental setup. And sorting strategy of brain microglia (CD45intCD11b+) by FACS. d. Western blot of MFE-2 in microglia from 8-month-old 5xFAD and untreated mice brain isolated by FACS. (Each sample contained a pool of microglia from 3 brains. Four samples per group were analyzed. Both male and female mice were used.) e. Overview of conditional MFE-2 knockout mice (ΔMFE-2) and MFE-2 knockout 5xFAD mice (5xFADΔMFE-2) production. f. Western blot of MFE-2 in isolated microglia by FACS from 8-week-old Flox and ΔMFE-2 mice brain (n = 4. Each lane counting for protein extracted from microglia of 3 brains p < 0.05. Both male and female mice were used.) g. Results of Morris water maze assays in Fig. 1g. Time in targeted quadrant and total swimming length were calculated. h. Representative immunofluorescences result of microglia distribution and morphology at early stage of AD (2 - 3-month-old) in 5xFAD and 5xFADΔMFE-2 mouse models (n = 5 per each group, male.) Mean microglial branch length was calculated by averaging the total length of branches using Leica software. Microglia in red and nuclei in blue. Scale bar, 100 μm. Statistical analyses were performed by two-tailed Student’s t-test. All data were presented as mean values ± SEM. Specific p values are indicated in the figure.
