Journal: International Journal of Molecular Sciences
Article Title: Clec7a Signaling in Microglia Promotes Synapse Loss Associated with Tauopathy
doi: 10.3390/ijms26072888
Figure Lengend Snippet: Clec7a–MD2 signaling mediates synaptic loss in PS19 mice. ( A – C ) Representative immunoblots of MD2 in ventral hippocampal lysate from WT or PS19 mice at 3 months ( A ), 6 months ( B ), and 9 months ( C ). ( D ) Representative immunoblots of hippocampus samples from AD patients. ( E ) Endogenous Clec7a was co-immunoprecipitated with MD2. ( F ) Quantitation of MD2 levels shown in panel ( A – C ) ( n = 6 mice per group, unpaired Student’s t test). ( G ) Quantitation of MD2 amounts in ( D ) ( n = 3 biologically independent samples per group, unpaired Student’s t test). ( H ) High resolution confocal images and 3D reconstructions show the presence of MD2 (magenta) tagged PSD95 (red) within Iba1 + cells (green) in the DG regions of hippocampal tissue from Laminarin-administered PS19 mice. White arrows indicating PSD95+ MD2+ puncta in Iba1 + cell. Scale bars, 5 μm. ( I ) Quantitation of MD2-PSD95 colocalized puncta ( n = 6 mice per group, one-way ANOVA test). ( J ) Quantitation of MD2-PSD95 colocalized puncta in Iba1 + cell ( n = 6 mice per group, one-way ANOVA test). * p < 0.05; ** p < 0.01; *** p < 0.001. Data are presented as mean ± SD.
Article Snippet: For the immunoprecipitation of MD2, 1.5 μg of biotinylated rabbit anti-MD2 antibody (GTX85517, GeneTex) was incubated with 400–500 μg of protein sample overnight at 4 °C.
Techniques: Western Blot, Immunoprecipitation, Quantitation Assay
