Journal: Food and chemical toxicology : an international journal published for the British Industrial Biological Research Association

Article Title: Inhibition of sEH via stabilizing the level of EETs alleviated Alzheimer's disease through GSK3β signaling pathway.

doi: 10.1016/j.fct.2021.112516

Figure Lengend Snippet: Fig. 4. Inhibition of sEH by TPPU alleviated neuroinflammation in Aβ-induced AD mice. (A–D) Representative immunohistochemistry images for GFAP in the dorsal (A), CA1 (B), CA3 (C), and DG (D) of the hippocampus. (E) Representative immunohistochemistry images for IBA-1 in the dorsal, CA1, CA3, and DG of the hip pocampus. (F) Inhibition of sEH by TPPU suppressed NF-κB signaling pathway involved in COX-2, Iba-1, IL-6, GFAP, p-p65, p65, p-IKKβ, and IKKβ. (G) Quantitative data of COX-2, Iba-1, IL-6, GFAP, p-p65/p65, and p-IKKβ/IKKβ. Data represent mean ± SEM, n = 3.

Article Snippet: The primary antibodies GAPDH, Aβ, Bax, IL-6, and GFAP were purchased from Proteintech (Wuhan, China).

Techniques: Inhibition, Immunohistochemistry

Journal: Current protocols

Article Title: Human Umbilical Cord Mesenchymal Stem Cell-Based in vitro Model for Neurotoxicity Testing.

doi: 10.1002/cpz1.423

Figure Lengend Snippet: Figure 6 Morphological and immunofluorescence analyses. (Top) Morphological examination of hMSCs and their neuronal differentiation at days 3 and 8 by phase-contrast microscopy. hMSCs exhibit the characteristic spindle-shaped morphology and form a homogeneous monolayer, typical of mesenchymal stem cells; in these hMSCs, no changes into a neuronal phenotype are observed. On the contrary, after transdifferentiation, cells adopt a neural-like morphology, with retractile cell bodies and long branching processes.On day 8, the morphological changes became more evident. (Bottom) The fluorescence signals of GFAP (green fluorescence), nestin (red fluorescence), and SOX-2 (green fluorescence) are well visible in hMSCs, and then decrease during the transdifferen- tiation process into hNLCs (3 and 8 days). Weak labeling of β-Tub III (green fluorescence), MAP-2 (green fluorescence), and NSE (red fluorescence) is observed on undifferentiated hMSCs and, on the contrary, increases in fluorescence intensity of these three neuronal markers are detected on hNLCs from days 3 to 8. Scale bar: 100 μm.

Article Snippet: Materials 1 mg/ml human fibronectin solution (PromoCell, cat. no. C43060) hMSCs (from Basic Protocol 1) Mesenchymal stem cell growth medium 2, ready-to-use (MSC medium; see recipe) Complete mesenchymal stem cell neurogenic differentiation medium (Ready-to-use) (neurogenic medium; see recipe) Test drug/chemical of interest Trypan blue solution 0.4% (Corning, cat. no. 25-900-CI) MTT ((3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) (Sigma-Aldrich, cat. no. M-2128) ATP reagent (CellTiter-Glo® 3D reagent, Promega, cat. no. G9681) Phosphate-buffered saline (1× PBS), without calcium, without magnesium (Carlo Erba Reagents, cat. no. LJ67802AP) 4% paraformaldehyde (PF; see recipe) Triton X-100 (Sigma-Aldrich, cat no. X100) Dry milk (BioRad, cat. no. 170-6404) Primary antibodies: Anti-human NSE conjugated to Alexa-Fluor® 594 (Santa Cruz Biotechnology, cat. no. sc-271384-AF594) Anti-human β-Tub III conjugated to Alexa-Fluor® 488 (Merck, cat. no. AB15708A4) Anti-human MAP-2 conjugated to Alexa-Fluor® 488 (Merck, cat. no. MAB3418X) Anti-human nestin conjugated to Cy3 (Merck, cat. no. MAB5326C3) Anti-human SOX2 conjugated to Alexa-Fluor® 488 (BioLegend, cat. no. 656110) Anti-human GFAP (Santa Cruz Biotechnology, cat. no sc-33673) Secondary antibody: Goat anti-Mouse IgG2b conjugated to Alexa-Fluor® 488 (Invitrogen, cat. no. A-21141) Hoechst 33258 (Invitrogen, cat. no. H3569) Fluoroshield (Sigma-Aldrich, cat. no. F6182) Laminar flow hood (Gelaire, BSB 4 - S, Class II) 6-well plates (SPL, cat. no. 30006) 96-well plates (SPL, cat. no. 330035)Coccini et al. 16 of 37 Current Protocols Figure 5 Schematic representation and timing of the steps described in Basic Protocol 2.

Techniques: Microscopy, Labeling