Journal: Cell Metabolism

Article Title: Regulation of Mother-to-Offspring Transmission of mtDNA Heteroplasmy

doi: 10.1016/j.cmet.2019.09.007

Figure Lengend Snippet:

Article Snippet: Mouse IgG (H+L) DyLight 800 , Rockland , 610-145-002.

Techniques: Recombinant, Modification, CyQUANT Assay, Proliferation Assay, Blocking Assay, Plasmid Preparation, Software

Journal: Cell stem cell

Article Title: Human uterine natural killer cells regulate differentiation of extravillous trophoblast early in pregnancy.

doi: 10.1016/j.stem.2023.12.013

Figure Lengend Snippet: Figure 1. Identification of uNK cell-restricted cytokines that have receptors on EVTs (A) Diagram showing key cell types in the first trimester placenta, with interactions between maternal uNK cells and fetal EVTs shown. VCT, villous cytotrophoblast; SCT, syncytiotrophoblast; GC, giant cell; iEVT, interstitial EVT; eEVT, endovascular EVTs; PV, perivascular cells; endo, endothelial cells; macro, macrophages. (B) FACS gating strategy for isolating KIR2DS1 single positive (sp), KIR2DL1sp, and KIR2DS1/KIR2DL1 double positive (dp) uNK cells after co-culture with 221-target cells expressing Cw*0501 (C2+HLA-C) or Cw*0802 (C1+HLA-C). uNK cells were gated as Live/CD56+CD3CD14CD19CD9+CD49+ cells. (C) Heatmap showing the log2-transformed fold change between culture with 221-C2+HLA-C and that with 221-C1+HLA-C targets across the three donors in each uNK cell subset for protein-coding genes specifically upregulated in the KIR2DS1sp subset (n = 3 decidua; false discovery rate [FDR] < 0.05, and fold change > 1.5). (D) Uniform manifold approximation and projection (UMAP) visualizations of the expression of the four uNK cell-restricted cytokines in different cell populations at the maternal-fetal interface based on our previous scRNA-seq data. dM, decidual macrophages; dS, decidual stromal cells; Endo, endothelial cells; Epi, epithelial glandular cells; dP, perivascular cells; DC, dendritic cells; fFB, fetal fibroblasts; HB, Hofbauer cells; ILC, innate lymphocyte cells; p, proliferative. (E) Intracellular staining of KIR2DS1sp uNK cells for cytokines XCL1 and CSF2 after 5 h co-culture with 221-C2+HLA-C compared with 221-C1+HLA-C targets (percentage of positive staining cells in KIR2DS1sp subset compared to isotype-matched control, n = 6). p values were obtained from one-sided paired Student t test.

Article Snippet: To stain for KIR2DL1 or KIR2DS1 in the same sample, cells were stained with 3 ml KIR2DL1 APC antibody (clone REA284; Miltenyi Biotec) in the cocktail with the other antibodies for 23 min. Then 7 ml KIR2DL1/S1 (clone 11PB6; Miltenyi Biotec) was added for the final 12 min. After washing in FACS buffer, cells were fixed in 2% paraformaldehyde (Alfa Aesar, J61899).

Techniques: Co-Culture Assay, Expressing, Transformation Assay, Staining, Control

Journal: Food Science & Nutrition

Article Title: Seahorse Attenuated DSS ‐Induced Depression in Mice by Inhibiting Neuroinflammation and Ferroptosis

doi: 10.1002/fsn3.70482

Figure Lengend Snippet: Effect of seahorses on DSS‐induced neuroinflammation in the hippocampal CA1 region ( n = 6). (A) Quantitative and morphological analysis of microglia in the whole hippocampus of the mouse hippocampus. (B) Immunofluorescence staining for NLRP3. (C) Immunofluorescence staining for TLR4. (D) Immunofluorescence staining for phosphorylated NF‐κB. Immunofluorescence staining for proinflammatory cytokines (E) IL‐1β, (F) IL‐6, and (G) TNF‐α. Scale bar = 200 μm. # p < 0.05 and ## p < 0.01 vs. control group; * p < 0.05 and ** p < 0.01 vs. DSS‐induced model group.

Article Snippet: Primary antibodies were against the following proteins: NF‐kB p65 (1:200; ab86299, Abcam), AIF (1:200; PA5143222, Thermo Fisher), IL‐6 (1:200; ab9324, Abcam), NLRP3 (1:200; ab263899, Abcam), TNF‐α (1:200; AF‐A10‐SP, R&D), GPX4 (1:200; NBP3‐07344, Novus Biologicals), BDNF (1:200; CST47808 , Cell Signaling Technology), SLC7A11 (1:200; MA5‐44922, Thermo Fisher), Neun (1:200; ab279297, Abcam), DCX (1:200; CST47808 , Cell Signaling Technology), Hexokinase (1:200; NBP1‐51644, Novus Biologicals), IBA1 (1:200; ab5076, Abcam), pNrf2 (1:200; PA5‐67520, Thermo Fisher), IL‐1β (1:200; cat log no. AF‐401‐SP, R&D), Claudin‐1 (1:200; PA5‐32350, Thermo Fisher), TLR4 (1:200; MAB2759‐SP, R&D).

Techniques: Immunofluorescence, Staining, Control