Journal: Frontiers in Pharmacology

Article Title: A protein–miRNA biomic analysis approach to explore neuroprotective potential of nobiletin in human neural progenitor cells (hNPCs)

doi: 10.3389/fphar.2024.1343569

Figure Lengend Snippet: (A–C) Phase-contrast images of neural progenitor cells were taken at a magnification of (A) ×40 and (B) ×100 captured using a Nikon Eclipse Ti-S inverted microscope. (C) Characterization of hiPSC-derived NPCs via flow cytometry.

Article Snippet: The iPSC-derived NPCs underwent flow cytometry analysis through a BD FACSCanto-II flow cytometry apparatus (Becton Dickinson, San Jose, CA, USA).

Techniques: Inverted Microscopy, Derivative Assay, Flow Cytometry

Journal: STAR Protocols

Article Title: Using the dCas9-KRAB system to repress gene expression in hiPSC-derived NGN2 neurons

doi: 10.1016/j.xpro.2021.100580

Figure Lengend Snippet:

Article Snippet: Two stable hiPSC-derived neuronal progenitor cells (hiPSC-NPCs) expressing dCas9 -KRAB (Addgene plasmid #99372) , Brennand Lab at Icahn School of Medicine at Mount Sinai , 553-S1-1 KRAB and 2607-1-4 KRAB.

Techniques: Recombinant, Purification, Virus, Titration, Saline, Lysis, Extraction, Western Blot, Plasmid Preparation, SYBR Green Assay, Bradford Protein Assay, Knockdown, RNA Sequencing, Gene Expression, Expressing, Software, CRISPR, Imaging, Microscopy, Real-time Polymerase Chain Reaction

Journal: Journal of Neurochemistry

Article Title: Altered metabolic function induced by Aβ‐oligomers and PSEN1 mutations in iPSC ‐derived astrocytes

doi: 10.1111/jnc.16267

Figure Lengend Snippet: Astrocytes process APP differentially in ‘healthy’ control versus fAD patient‐derived cells. Control and fAD (PSEN1 mutation)‐derived astrocytes (Day 45) were characterised using immunofluorescence and APP processing was assessed using ELISA and ADAM 10 activity. (a) Representative images of fAD (PSEN1 mutation) derived astrocytes. Cells were stained using immunofluorescent antibodies for astrocytic markers ALDH1L1 (green), S100β (Green) and GFAP (red). nuclei were counterstained with DAPI (blue). Scale bars: 100 μM. (b) Characterisation of ADAM10 enzymatic activity (RFU) and (c) soluble APPα (ng/mL) in control and PSEN1 (A246E, L268V and R278I). Pooled PSEN1 samples compared to control are displayed. (d) Aβ 1–40 (pg/mL), (e) Aβ 1–42 (pg/mL), (f) Aβ42/40 ratio, (g) aggregated Aβ (pg/mL) were measure in control and fAD patient‐derived astrocytes after 48 h. Pooled PSEN1 samples compared to control are displayed. Results are expressed as ± SD, n = 3 p < 0.05 (*), p < 0.01 (**), p < 0.001 (***). For direct comparison between control and PSEN1, unpaired t ‐tests were performed. Comparisons between individual PSEN1 lines were performed using ANOVA followed by Dunnet's post‐test. Each replicate or ‘ n ’ represents an independent culture preparation and is displayed as an individual data point.

Article Snippet: Healthy control (ax0018) and PSEN1 human neural precursor cells (hNPCs) carrying L286V (ax0112) and A246E (ax0114) mutations were purchased from Axol Bioscience (Cambridge, UK).

Techniques: Control, Derivative Assay, Mutagenesis, Immunofluorescence, Enzyme-linked Immunosorbent Assay, Activity Assay, Staining, Comparison

Journal: Journal of Neurochemistry

Article Title: Altered metabolic function induced by Aβ‐oligomers and PSEN1 mutations in iPSC ‐derived astrocytes

doi: 10.1111/jnc.16267

Figure Lengend Snippet: fAD astrocytes display markers of gliosis compared to ‘healthy’ control astrocytes. Cytokine, GFAP, 8‐isoprostane levels in media or cellular lysates were measured in control and fAD astrocytes (45 days old). (a) Levels of GFAP were measure in the cell lysates or (b) cell culture media using ELISA (ng/mL). (c) Isoprostane levels were also measured in cellular lysates (pg/mg). Pooled control and fAD cell samples were compared in (d) cellular GFAP and (e) secreted GFAP, (f) isoprostanes. Fold change in the accumulation of (g) IL‐6 and (h) IL8 following protein transport inhibition measured using flow cytometry. Results are expressed as ± SD, n = 3 p < 0.05 (*), p < 0.01 (**), p < 0.001 (***). For direct comparison between control and PSEN1, unpaired t‐tests were performed. Comparisons between individual PSEN1 lines were performed using ANOVA followed by Dunnet's post‐test. Each replicate or ‘ n ’ represents an independent culture preparation and is displayed as an individual data point.

Article Snippet: Healthy control (ax0018) and PSEN1 human neural precursor cells (hNPCs) carrying L286V (ax0112) and A246E (ax0114) mutations were purchased from Axol Bioscience (Cambridge, UK).

Techniques: Control, Cell Culture, Enzyme-linked Immunosorbent Assay, Inhibition, Flow Cytometry, Comparison

Journal: Journal of Neurochemistry

Article Title: Altered metabolic function induced by Aβ‐oligomers and PSEN1 mutations in iPSC ‐derived astrocytes

doi: 10.1111/jnc.16267

Figure Lengend Snippet: fAD‐derived astrocytes display significantly altered metabolic profiles compared to controls. (a) Heatmap displaying differential compounds identified using metabolomic analysis. Summary of pathway analysis for the comparison of control and b) A246E, (c) L286V and (d) R278 fAD patient‐derived astrocytes. The pathway analysis results of PSEN1 astrocytes compared with control. The colour graduated from white to and red indicates the degree of significance, the size of bubble represents the number of metabolites hit in the pathway. (e) Bar charts indicating intensity changes in key metabolites represented within metabolic pathway that were detected in astrocytes carrying PSEN1 mutations. Data shown are expressed as ± SD, n = 6. Each replicate or ‘ n ’ represents an independent culture preparation.

Article Snippet: Healthy control (ax0018) and PSEN1 human neural precursor cells (hNPCs) carrying L286V (ax0112) and A246E (ax0114) mutations were purchased from Axol Bioscience (Cambridge, UK).

Techniques: Derivative Assay, Comparison, Control