Journal: Viruses

Article Title: EcoHIV-Infected Mice Show No Signs of Platelet Activation

doi: 10.3390/v16010055

Figure Lengend Snippet: Synthesis of EcoHIV in the spleen peaked after 7 days but then became latent. Mice were infected with EcoHIV (i.v.) and euthanized either after 1, 2, 7, or 42 days. Spleen RNA was isolated, and 500 ng was used for RT-qPCR. ( A ) EcoHIV gag transcript is present in the spleen of infected mice, and its synthesis peaked after 7 days of infection before going into the latent phase on day 42. ( B ) EcoHIV spliced vif is present in the spleen of infected mice, and its synthesis peaked after 7 days of infection before going into the latent phase on day 42. The presence of gag and spliced vif indicates de novo virus synthesis and processing. ( C ) Anti-Gag antibodies in mouse plasma (uninfected and 42 days post infection) ( left panel). Human plasma samples (healthy controls and HIV-1-naïve patients) were used as negative and positive controls ( right panel). ( D ) Representative images of uninfected and infected mouse (42 days) brains stained for CD68 + microglia. There is a clear activation of the microglial in the brain (black arrows), which is consistent with previously reported data. Each data point represents a mouse. Data graphed as mean ± SEM. Statistical analyses were performed using one-way ANOVA or unpaired t -test. * p < 0.05 **, p < 0.01, *** p < 0.001, and **** p < 0.0001.

Article Snippet: Overnight incubation was carried out at 4 °C in a 1:2000 dilution of a primary antibody (Rat anti-mouse CD68, MCA1957-BioRad, CA, USA), followed by 1 h incubation at room temperature in a 1:1000 dilution of a secondary antibody (biotinylated Donkey anti-Rat IgG (712-065-153, Jackson ImmunoResearch, PA, USA).

Techniques: Infection, Isolation, Quantitative RT-PCR, Virus, Clinical Proteomics, Staining, Activation Assay

Journal: bioRxiv

Article Title: Unique Microglial Transcriptomic Signature within the Hippocampal Neurogenic Niche

doi: 10.1101/2021.08.29.457876

Figure Lengend Snippet: (A) Volcano plot showing differentially expressed genes in SGZ microglia compared to other hippocampal microglia. Statistically significant genes (up or down-regulated) are represented by red dots (LFC > 0.25 and p-val < 10e-32). (B) Violin Plots showing comparison of expression profiles of key genes in SGZ cluster (cluster 8) vs homeostatic clusters (13 and 14). (C) Gene set enrichment analysis of upregulated genes in cluster 8 showing select ontology related to neurogenesis. (D) Immunohistochemistry staining for Cd9 colocalized with Iba1 in the dentate gyrus. Decreased immunoreactivity to Cd9 as indicated with an arrow in molecular layer (middle) with increased Iba1 colocalization in SGZ as indicated by arrowheads (right panel). (E) Immunohistochemistry staining for Tmem119 colocalized with Iba1 in the dentate gyrus. Decreased Tmem119 immunoreactivity in SGZ (arrow in middle panel) and sparse colocalization with Iba1 in SGZ indicated by arrowheads (right panel). Scale for D and E = 80 µm.

Article Snippet: Samples were then incubated overnight in primary antibody at the following concentrations 1:500 Iba1 (Wako 019-19741), 1:100 Cd68 (Biorad MCA1957), 1:200 Cd9 (BioLegend 124802) at 4%C.

Techniques: Expressing, Immunohistochemistry, Staining

Journal: medRxiv

Article Title: Phospholipase D3 contributes to Alzheimer’s disease risk via disruption of Aβ clearance and microglia response to amyloid plaques

doi: 10.1101/2022.01.31.22270175

Figure Lengend Snippet: A. Representative images of APP/PS1x Pld3 -WT and APP/PS1x Pld3 -KO mice co-stained with Iba1 (total microglia), CD68 (activated microglia), and X-34 (β-sheet-rich dense cores). APP/PS1x Pld3 -WT (n=20), APP/PS1x Pld3 -KO (n=19). B. Quantification of Iba1 localization within 15 µm of the X-34+ dense core plaques (*, p=0.03). C. Quantification of CD68 localization within 15 µm of the dense plaques. D. Quantification of Iba1 and CD68 colocalization. Graphs represent mean ± SEM. Analyzed by two-tailed Student’s t test with a ROUT outlier analysis (Q=1%).

Article Snippet: X-34-stained samples were immunostained with antibodies HJ3.4 (2.4ug/mL; α-Aβ1-13) or Iba1 (Abcam ab5076) and CD68 (BioRad MCA1957) at 4°C overnight.

Techniques: Staining, Two Tailed Test