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: bioRxiv

Article Title: Proteomic characterization of neuronal extracellular vesicle interactomes in Alzheimer’s disease mouse model through TurboID-based proximity labeling

doi: 10.1101/2025.10.22.684016

Figure Lengend Snippet: (A) Schematic overview of PLA validation and in silico modeling workflows of CD9 interactions with of proteins of interest. (B) PLA conjugation of Ncam1 with streptavidin in the TB and SB groups in both WT and APP NLGF mice where mCherry represents TurboID infected neuron, blue represents DAPI and green represents PLA conjugation. (C) Comparison of PLA conjugation (surface count) of Ncam1 with streptavidin between all groups (One-way ANOVA). (D) HADDOCK modeling of CD9 protein-protein docking with Ncam1 where zoomed in images highlight specific residue interactions. (E) PLA conjugation of Gabrb3 with streptavidin in the TB and SB groups in both WT and APP NLGF mice. (F) Comparison of PLA conjugation (surface count) of Gabrb3 with streptavidin between all groups (One-way ANOVA). (G) HADDOCK modeling of CD9 protein-protein docking with Gabrb3 where zoomed in images highlight specific residue interactions. (H) PLA conjugation of Gad1 with streptavidin in the TB and SB groups in both WT and APP NLGF mice. (I) Comparison of PLA conjugation (surface count) of Gad1 with streptavidin between all groups (One-way ANOVA). (J) HADDOCK modeling of CD9 protein-protein docking with Gad1 where zoomed in images highlight specific residue interactions.

Article Snippet: Following primary antibody incubation, sections were washed and incubated with Alexa Fluor 488-conjugated secondary antibodies, along with Alexa Fluor 647-conjugated streptavidin (016-600-084, Jackson Immunoresearch) to detect biotinylated proteins.

Techniques: Biomarker Discovery, In Silico, Conjugation Assay, Infection, Comparison, Residue