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Fig. 5 | Acta Neuropathologica Communications

Fig. 5

From: Regulating microglial miR-155 transcriptional phenotype alleviates Alzheimer’s-induced retinal vasculopathy by limiting Clec7a/Galectin-3+ neurodegenerative microglia

Fig. 5

Targeting microglial miR-155 protected retinal blood vessels and reduced vascular amyloidosis in APP/PS1 mouse retinas. a Representative images of immunofluorescent staining for 4G8 of Aβ (magenta), Claudin-1 (red), lectin for blood vessels (green) and DAPI (blue) on isolated retinal blood vessels from all four genotypes—WT, WT:miR-155cKO, APP/PS1, and APP/PS1:miR-155cKO—in 8-month-old mice. Images were obtained using the 63 × microscope objective. Arrows indicate vascular Aβ. Scale bars = 10 µm. b Quantitative analysis of Claudin-1-immunoreactivity (IR) in isolated retinal blood vessels from all 8-month-old experimental groups (n = 24 total, n = 6 each group). c Densitometric analysis of western blotting protein bands of Claudin-1 normalized by β-actin control for retinal lysates from all experimental groups (n = 48 total, n = 6 each group). d Representative images of immunofluorescent staining for 11A50-B10 of Aβ40 (magenta), ZO-1 (red), lectin for blood vessels (green) and DAPI (blue) on isolated retinal blood vessels from all four genotypes—WT, WT:miR-155cKO, APP/PS1, and APP/PS1:miR-155cKO—in 8-month-old mice. Images were obtained using the 63 × microscope objective. Arrows indicate vascular Aβ. Scale bars = 10 µm. e Quantitative analysis of ZO-1-IR in isolated retinal blood vessels from same mouse cohort as shown in panel b. f Quantitative analysis of 11A50-B10 for Aβ40-IR in isolated retinal blood vessels from 8-month-old APP/PS1 experimental groups (n = 12 total, n = 6 each group). g Representative images of immunofluorescent staining of 4G8 for Aβ (magenta) on isolated retinal blood vessels from 8-month-old APP/PS1 or APP/PS1:miR-155cKO mice groups. h Quantitative analysis of 4G8 for Aβ in isolated retinal blood vessels from the same mouse cohort shown in panel f. i Densitometric analysis of western blotting protein bands of MMP-9 normalized by β-actin control for retinal lysates from APP/PS1:miR-155cKO vs. APP/PS1 mice (n = 12 total, n = 6 each group). j, k Pearson’s coefficient (r) correlation between j. retinal vascular Aβ40 and retinal Clec7a+ microglia, k retinal vascular Claudin-1-IR and retinal Clec7a+ microglia or retinal vascular Aβ-IR. Data from individual mice (circles) as well as group means ± SEMs are shown. Black-filled circles represent male and clear circles represent female animals. *p < 0.05, **p < 0.01, by two-way or three-way ANOVA with Tukey’s post-hoc multiple comparison test. Two group statistical analysis was performed using an unpaired two-tailed Student t-test. Fold changes and percentage decreases are shown in red

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