Figure 1.
Conditional deletion of Atg7 in microglia impairs their response to Aβ plaques. (A) Immunoblotting of microglial lysates for ATG7 and LC3I/II. Actin was assessed as a loading control. Microglia were sorted from 5-mo-old Atg7fl/fl- and Atg7ΔMG-5xFAD mice. Experiments were repeated three times. (B) Representative TEM images of microglia (CD45lo CD11b+) sorted from 5-mo-old Atg7fl/fl- and Atg7ΔMG-5xFAD mice. Multivesicular structures are indicated by white arrows. Scale bar = 500 nm. (C) Quantification of the number of multivesicular structures per microglia observed in TEM images. Total of 30 cells from each genotype was analyzed. Each point represents one cell. *, P < 0.05 by two-tailed unpaired t test. Data are presented as mean ± SEM. (D) Representative confocal images from the cortex of Atg7fl/fl- and Atg7ΔMG-5xFAD mice at 5 and 12 mo of age, showing Methoxy-X04–labeled Aβ plaques (blue), IBA1+ microglia (green), and PU.1 (red). Scale bar = 100 μm. (E) Quantification of microglia density (microglia number per mm2) in Atg7fl/fl- and Atg7ΔMG-5xFAD mice. Atg7fl/fl-5xFAD (5-mo-old), n = 7; Atg7ΔMG-5xFAD (5-mo-old), n = 6; Atg7fl/fl-5xFAD (12-mo-old), n = 4; and Atg7ΔMG-5xFAD (12-mo-old), n = 9. Each point represents data from one mouse with two technical repeats. *, P < 0.05; **, P < 0.01 by two-tailed unpaired t test. Data are presented as mean ± SEM. (F and G) Quantification of the average number of plaque-associated microglia (microglia within 15-μm radius from plaque surfaces). Each point represents the average of two technical repeats from one mouse. ***, P < 0.001; ****, P < 0.0001 by two-tailed unpaired t test. Data are presented as mean ± SEM. (H and I) Quantification of the number of non-plaque–associated microglia (microglia that are 15 μm away from plaque surfaces). Each point represents the average of two technical repeats from one mouse. *, P < 0.05; ***, P < 0.001 by two-tailed unpaired t test. Data are presented as mean ± SEM. (J) Representative confocal images of Methoxy-X04 (blue), IBA1 (green), and CD11c (red) staining in the cortex of 5-mo-old Atg7fl/fl- and Atg7ΔMG-5xFAD mice. Scale bar = 100 μm. (K) Quantification of the ratio of the IBA1+CD11c+ volume to IBA1+ volume in confocal images, which represents the percentage of CD11c+ microglia. Each point represents the average of four technical repeats from one mouse. **, P < 0.01 by two-tailed unpaired t test. Data are presented as mean ± SEM. Source data are available for this figure: SourceData F1. Refer to the image caption for details.

Conditional deletion of Atg7 in microglia impairs their response to Aβ plaques. (A) Immunoblotting of microglial lysates for ATG7 and LC3I/II. Actin was assessed as a loading control. Microglia were sorted from 5-mo-old Atg7fl/fl- and Atg7ΔMG-5xFAD mice. Experiments were repeated three times. (B) Representative TEM images of microglia (CD45lo CD11b+) sorted from 5-mo-old Atg7fl/fl- and Atg7ΔMG-5xFAD mice. Multivesicular structures are indicated by white arrows. Scale bar = 500 nm. (C) Quantification of the number of multivesicular structures per microglia observed in TEM images. Total of 30 cells from each genotype was analyzed. Each point represents one cell. *, P < 0.05 by two-tailed unpaired t test. Data are presented as mean ± SEM. (D) Representative confocal images from the cortex of Atg7fl/fl- and Atg7ΔMG-5xFAD mice at 5 and 12 mo of age, showing Methoxy-X04–labeled Aβ plaques (blue), IBA1+ microglia (green), and PU.1 (red). Scale bar = 100 μm. (E) Quantification of microglia density (microglia number per mm2) in Atg7fl/fl- and Atg7ΔMG-5xFAD mice. Atg7fl/fl-5xFAD (5-mo-old), n = 7; Atg7ΔMG-5xFAD (5-mo-old), n = 6; Atg7fl/fl-5xFAD (12-mo-old), n = 4; and Atg7ΔMG-5xFAD (12-mo-old), n = 9. Each point represents data from one mouse with two technical repeats. *, P < 0.05; **, P < 0.01 by two-tailed unpaired t test. Data are presented as mean ± SEM. (F and G) Quantification of the average number of plaque-associated microglia (microglia within 15-μm radius from plaque surfaces). Each point represents the average of two technical repeats from one mouse. ***, P < 0.001; ****, P < 0.0001 by two-tailed unpaired t test. Data are presented as mean ± SEM. (H and I) Quantification of the number of non-plaque–associated microglia (microglia that are 15 μm away from plaque surfaces). Each point represents the average of two technical repeats from one mouse. *, P < 0.05; ***, P < 0.001 by two-tailed unpaired t test. Data are presented as mean ± SEM. (J) Representative confocal images of Methoxy-X04 (blue), IBA1 (green), and CD11c (red) staining in the cortex of 5-mo-old Atg7fl/fl- and Atg7ΔMG-5xFAD mice. Scale bar = 100 μm. (K) Quantification of the ratio of the IBA1+CD11c+ volume to IBA1+ volume in confocal images, which represents the percentage of CD11c+ microglia. Each point represents the average of four technical repeats from one mouse. **, P < 0.01 by two-tailed unpaired t test. Data are presented as mean ± SEM. Source data are available for this figure: SourceData F1.

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