Journal: Immunity
Article Title: Targeted complement inhibition at synapses prevents microglial synaptic engulfment and synapse loss in demyelinating disease
doi: 10.1016/j.immuni.2019.12.004
Figure Lengend Snippet: (A) Schematic of the course of clinical symptoms in EAE of C57Bl6/J WT mice. Mice were analyzed at the onset of moderate clinical symptoms, typically observed between day 10–12 post-immunization. (B,C) Representative images of the LGN from CFA-control and EAE-induced mice immunostained against (B) presynaptic VGluT2 (green) and postsynaptic PSD-95 (red) or (C) presynaptic VGluT1 (green) and postsynaptic Homer1 (red). (Bi-Ci) Quantification of synaptic staining in B,C. n=4 mice per condition from one experiment. (D) Schematic for tamoxifen-induced genetic depletion of mature oligodendrocytes in Plp1-CreERT;ROSA26-EGFP-DTA (DTA) mice. Tamoxifen-injected ROSA26-EGFP-DTA mice were used as controls (ctrl.). The LGN of mice was analyzed at 35 days post induction. (E,F) Representative confocal images of the LGN from control and DTA mice immunostained against (E) presynaptic VGluT2 (green) and postsynaptic PSD-95 (red) or (F) presynaptic VGluT1 (green) and postsynaptic Homer1 (red). (Ei-Fi) Quantification of synaptic staining in E,F. (E,F) n=5 ctrl./4 DTA mice from two experiments. (B-F) Scale bars, 5 μm. Data represent mean ± SEM, significant differences with **P < 0.01, ****P < 0.0001, t-test. See also Figs. S2,3,5.
Article Snippet: The following primary monoclonal (mAb) and polyclonal (pAb) antibodies have been used: mouse mAb α-ALDH1L1 (clone N103/39, Millipore, MABN495, 1:1000), mouse mAb α-APP (clone 22C11, Millipore, MAB348, 1:200), rabbit mAb α-C1q (clone 4.8, Abcam, ab182451, 1:100), rat mAb α-C3 (clone 11H-9, Abcam, ab11862, 1:500), rabbit pAb α-CASPR (provided by Matthew N. Rasband, 1:100), rabbit mAb α-CD3 (clone SP7, Abcam, ab16669, 1:100), mouse mAb α-CD8 (clone 8/114B, Thermo Fisher Scientific, MA5–13473, 1:20), rat mAb α-CD45 (clone IBL-3/16, BioRad, MCA1388, 1:100), mouse mAb α-CD68 (clone KP1, Abcam, ab955, 1:200), rat mAb α-CD68 (clone FA-11, AbD Serotec, MCA1957, 1:1000), rat mAb α-Clec7a (InvivoGen, mabg-mdect, 1:200), rabbit pAb α-Clec12a (LSBio, LS-C377776, 1:500), rabbit pAb α-cleaved caspase-3 (Cell Signaling Technologies, #9661, 1:200), mouse mAb α-Crry (clone TLD-1C11, Santa Cruz, sc-53530, 1:100), chicken pAb α-EGFP (Abcam, ab13970, 1:500), mouse mAb α-FoxP3 (clone 206D, BioLegend, 320102, 1:50), mouse mAb α-GFAP (clone G-A-5, Sigma, G3893, 1:500), rabbit pAb α-Homer1 (Synaptic Systems, #160003, 1:1000), rabbit pAb α-Iba1 (Wako Chemicals, #019–19741, 1:500), rat mAb α-LAMP2 (clone GL2A7, Abcam, ab13524, 1:200), rat mAb α-Ly6C (clone HK1.4, BioLegend, 128016, 1:300), mouse mAb α-MAG (clone 513, Millipore, MAB1567, 1:100), rat mAb α-MBP (clone 12, Millipore, MAB386, 1:500), mouse mAb α-MOG (clone 8–18C5, Millipore, MAB5680, 1:200), rabbit pAb α-Neurofilament 200 (Sigma, N4142, 1:1000), chicken pAb α-NeuN (Millipore, ABN91, 1:1000), rabbit pAb α-P2RY12 (AnaSpec, AS-55043A, 1:2000), rat mAb α-P2RY12 (clone S16007D, BioLegend, 848002, 1:100), mouse mAb α-PSD-95 (clone 6G6–1C9, Millipore, MAB1596, 1:100), guinea pig pAb α-RBPMS (PhosphoSolutions, 1832-RBPMS, 1:500), rabbit pAb α-βIV-spectrin (provided by Matthew N. Rasband, 1:100), guinea pig pAb α-VGluT1 (Millipore, ab5905, 1:2000), and guinea pig pAb α-VGluT2 (Millipore, ab2251, 1:2000).
Techniques: Staining, Injection
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