Journal: Journal of Lipid Research

Article Title: The cholesteryl ester transfer protein (CETP) raises cholesterol levels in the brain

doi: 10.1016/j.jlr.2022.100260

Figure Lengend Snippet: CETP promotes ABCA7 and TREM2 expression in the liver. A: Feeding schedule and study design. Wt and CETP tg mice were fed for 3 months starting at the age of 2 months. Biochemical analyses were performed at 5 months. B–D: Normalized RT-qPCR from mouse liver tissue. B: H mgcr , C: L dlr , and D: L rp 1 expression, n = 6–14, mean ± SEM; 2-way ANOVA, Tukey’s multiple comparison test. E: Western blot analysis of ABCA7 and TREM2 from liver lysates. Antibodies used: rabbit-anti-GAPDH (14C10; Cell Signaling), anti-TREM2 (Mab1729; R&D Systems), and anti-ABCA7 (polyclonal; Thermo Fisher Scientific), n = 6; mean ± SEM, Student's t -test. F–I: Expression analysis of ABCA7 and TREM2 from liver samples. Normalized RT-qPCR levels of liver F: A bca 7 and H: T rem 2 ; n = 6–14, mean ± SEM; 2-way ANOVA, Tukey’s multiple comparison test. Western blot quantification of G: ABCA7 and I: TREM2; n = 6; mean ± SEM, Student's t -test.

Article Snippet: Primary antibodies used were 22C11 (Millipore), anti-GAPDH (14C10; Cell Signaling), TP2 (kind gift of the Ottawa Heart Institute), anti-triggering receptor expressed in myeloid cells 2 (TREM2) (Mab1729; R&D Systems) and anti-ABCA7 (polyclonal; Thermo Fisher Scientific), and horseradish peroxidase-coupled secondary antibodies (Promega).

Techniques: Expressing, Quantitative RT-PCR, Comparison, Western Blot

Journal: Frontiers in Pharmacology

Article Title: Xixin Decoction’s novel mechanism for alleviating Alzheimer’s disease cognitive dysfunction by modulating amyloid-β transport across the blood–brain barrier to reduce neuroinflammation

doi: 10.3389/fphar.2024.1508726

Figure Lengend Snippet: XXD Upregulates TREM2 Expression and Downregulates IBA1, TLR1, and TLR2 Expression, Inhibiting Microglial Activation; (A) and (E) Analysis of relative IBA1 protein expression levels and average fluorescence intensity of IBA1 in the hippocampal CA1 region of mice; (B) and (F) Analysis of relative TLR1 protein expression levels and average fluorescence intensity of TLR1 in the hippocampal CA1 region of mice; (C) and (G) Analysis of relative TLR2 protein expression levels and average fluorescence intensity of TLR2 in the hippocampal CA1 region of mice; (D) and (H) Analysis of relative TREM2 protein expression levels and average fluorescence intensity of TREM2 in the hippocampal CA1 region of mice; (n = 6); Scale bar: 20μm; images captured at ×400 magnification; ** p < 0.01, * p < 0.05.

Article Snippet: Anti-TLR1 antibody (AA07135689), Anti-TLR2 antibody (BA06081077), and Anti-TREM2 antibody (BA08065558) were purchased from Bioss (Beijing, China).

Techniques: Expressing, Activation Assay, Fluorescence

Journal: Brain, behavior, and immunity

Article Title: Early life stress impairs synaptic pruning in the developing hippocampus

doi: 10.1016/j.bbi.2022.09.014

Figure Lengend Snippet: ELS reduces expression of TREM2 and CD11b while increasing CX3CR1 in P17 microglia. (A) Scatter plots for quantifying Trem2 positive microglia in CTL, LB and UPS. (B) Percentage of TREM2-positive microglia. Median fluorescence intensity (MFI) plots and surface expression of CD11b (C-D), CD45 (E-F) and CX3CR1 (G-H). CTL-males: n = 16, CTL-females: n = 14, LB males: n = 10, LB-females: n = 9, UPS-males: n = 12, UPS-females: n = 16. Error bars represent mean ± SEM. **p < 0.01, ***p < 0.001, ****p < 0.0001.

Article Snippet: Microglia were isolated from the hippocampus as described above and blocked with rat- anti CD16/CD32 Fc antibodies 1:100 (Cat# 553142, BD Biosciences) and then stained with rat-anti CD11b-PeCy7 1:100 (Cat# 25-0112-82, ebiosciences), rat-anti CD45-PeCPCy5.5 antibodies 1:100 (Cat# 45-0451-82, ebiosciences), APC-rat anti Trem2 antibodies 1:10 (Cat# FAB17291A R & D systems), or APC-mouse anti CX3CR1 antibodies 1:100 (Cat# 149008) for 25 min on ice.

Techniques: Expressing, Fluorescence

Journal: Brain, behavior, and immunity

Article Title: Early life stress impairs synaptic pruning in the developing hippocampus

doi: 10.1016/j.bbi.2022.09.014

Figure Lengend Snippet: Trem2 is essential for normal phagocytic activity. (A-B) Effects of Trem2 genotype (WT, Hets, Ko) on ex vivo phagocytic activity in microglia isolated from the hippocampus of 17-day old pups (n = 10–17 pups per group, 50 % females). (C) Representative confocal images (top row) and Imaris models (middle and lower rows) of microglia from Trem2 wildtype (WT), heterozygous (Hets) and knockout P17 littermates. Staining for Iba1 (green), CD68 (blue), and PSD95 (red). Middle row: reconstruction of Iba1 & CD68 staining. Lower row: reconstruction of CD68 and PSD95 staining inside microglia. Effects of Trem2 genotype on microglial volume (D), CD68 volume inside microglia (E), number of PSD95 puncta in microglia (F) and PSD95 puncta inside CD68 (G), n = 5 cells per mouse and 4–5 mice per group, 50 % females). Error bars represent mean ± SEM. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001.

Article Snippet: Microglia were isolated from the hippocampus as described above and blocked with rat- anti CD16/CD32 Fc antibodies 1:100 (Cat# 553142, BD Biosciences) and then stained with rat-anti CD11b-PeCy7 1:100 (Cat# 25-0112-82, ebiosciences), rat-anti CD45-PeCPCy5.5 antibodies 1:100 (Cat# 45-0451-82, ebiosciences), APC-rat anti Trem2 antibodies 1:10 (Cat# FAB17291A R & D systems), or APC-mouse anti CX3CR1 antibodies 1:100 (Cat# 149008) for 25 min on ice.

Techniques: Activity Assay, Ex Vivo, Isolation, Knock-Out, Staining