Journal: Current Research in Toxicology

Article Title: Neural organoids incorporating microglia to assess neuroinflammation and toxicities induced by known developmental neurotoxins ☆

doi: 10.1016/j.crtox.2025.100252

Figure Lengend Snippet: Generation and immunofluorescence characterization of planar neuroimmune organoid. Organoids are generated in 96-well plates by the addition of iPSC-derived cells to the top of a 1) PEG-based hydrogel. Neuroimmune organoids were created by sequentially adding 2) neural progenitor cells (NPCs), 3) endothelial cells (ECs) and mesenchymal cells (MSCs), and finally 4) microglia (MGs). Organoids are cultured 21–28 days from initial NPC plating prior to treatments. For the neurotoxicity assessment herein, organoids were treated on day 23 and dosing occurred for 4 days. Cell culture supernatants were collected each day. On day 4, organoids were either harvested for transcriptional analysis or fixed for immunocytochemistry analysis. Representative images show the glial components of the neuroimmune organoids. Astrocytes and microglia were stained using antibodies to detect GFAP (green, left) or IBA1 (orange, right), respectively. Neuronal staining was performed using β3 tubulin (blue, both panels). The images shown are maximum intensity projections of 20X confocal images.

Article Snippet: Donkey anti-rabbit AF555 antibody (1:500, Thermo A31572), and AF-647 conjugated anti β3 tubulin antibody (1:250, RnD Systems IC1195R), was incubated overnight at 4 °C and washed three times in PBS.

Techniques: Immunofluorescence, Generated, Derivative Assay, Cell Culture, Immunocytochemistry, Staining

Journal: Biology

Article Title: sTREM2 Differentially Affects Cytokine Expression in Myeloid-Derived Cell Models via MAPK-JNK Signaling Pathway.

doi: 10.3390/biology13020087

Figure Lengend Snippet: Figure 3. The effects of a TREM2 antibody (mAb) on sTREM2-induced cytokine expression: THP-1 cells were treated with 0.1 µg/mL sTREM2, mAb, or both sTREM2+mAb for 2, 4, 6, and 8 h, and cytokine expression of (A) TNF-α, (B) IL-1β, (C) TARC, and (D) IL-10 was assessed using qPCR. Data presented as Mean + SD.

Article Snippet: A commercially available rat anti-human IgG2B TREM2 antibody (R&D Systems) was used to assess the antibody’s ability to bind and neutralize sTREM2.

Techniques: Expressing

Journal: Biology

Article Title: sTREM2 Differentially Affects Cytokine Expression in Myeloid-Derived Cell Models via MAPK-JNK Signaling Pathway.

doi: 10.3390/biology13020087

Figure Lengend Snippet: Figure 4. Inhibition of NLRP-3 inflammasome-mediated release of IL-1β induced by the complex of sTREM2+mAb. THP-1 cells were treated for 6 h with 0.1 µg/mL sTREM2, TREM2 antibody (mAb), or 0.1 µg/mL sTREM2+mAb (a monoclonal TREM2 antibody) and 0.1 µg/mL sTREM2+mAb with a selective NLRP3 inhibitor MCC950 (7.5 nM). Data presented as Mean + SD. * p < 0.05; ** p < 0.01; *** p < 0.001.

Article Snippet: A commercially available rat anti-human IgG2B TREM2 antibody (R&D Systems) was used to assess the antibody’s ability to bind and neutralize sTREM2.

Techniques: Inhibition

Journal: Biology

Article Title: sTREM2 Differentially Affects Cytokine Expression in Myeloid-Derived Cell Models via MAPK-JNK Signaling Pathway.

doi: 10.3390/biology13020087

Figure Lengend Snippet: Figure 6. The effects of Syk, NFκB, and JNK inhibitors on sTREM2-induced cytokine expression (A). The effect of TREM2 inhibitor piceatannol (15 µM) on cytokine expression; (B). The effect of MG-132 (100 nM) on cytokine expression; (C). The effect of JNK inhibitor SP600125 (90 nM) on cytokine. Data presented as Mean + SD.

Article Snippet: A commercially available rat anti-human IgG2B TREM2 antibody (R&D Systems) was used to assess the antibody’s ability to bind and neutralize sTREM2.

Techniques: Expressing

Journal: Viruses

Article Title: Influence of Canonical and Non-Canonical IFNLR1 Isoform Expression on Interferon Lambda Signaling

doi: 10.3390/v15030632

Figure Lengend Snippet: FLAG-IFNLR1 isoforms and co-receptor IL10RB. Schematic depicting the structure of each FLAG-tagged IFNLR1 isoform and IL10RB. Box1 and Box2 depict the Jak1 binding domain that is fully present in FLAG-IFNLR1 isoform 1, truncated in FLAG-IFNLR1 isoform 2, and absent in FLAG-IFNLR1 isoform 3. Image created with BioRender.com using default settings to represent a transmembrane protein.

Article Snippet: We did not detect any differences in IL10RB surface expression by flow cytometry (FAB874G, R&D Systems) or any differences in cell viability between lines by cell counting or flow cytometry using a Zombie viability stain.

Techniques: Binding Assay