Journal: Molecular Neurodegeneration

Article Title: LRP1 is a neuronal receptor for α-synuclein uptake and spread

doi: 10.1186/s13024-022-00560-w

Figure Lengend Snippet: Generation and validation of human induced pluripotent stem cells (iPSCs)-derived neurons (iPSNs) with LRP1 gene knockout ( LRP1 -KO). a Schematic diagram of the workflow for LRP1 -KO iPSC generation, neural differentiation, and protein uptake assays. LRP1 -KO iPSC colonies were obtained using CRISPR/Cas9 gene editing stratagy. Neural progenitor cells (NPCs) were then induced from the iPSCs and further differentiated into iPSNs. On day 14 to 16 of iPSN differentiation, neurons were treated with fluorescently labeled proteins and the uptake was measured by flow cytometry. b Immunofluorescence images showing NPCs from all three lines (WT, LRP1 -KO#1, and LRP1 -KO#2) were positive for neural precursor marker, Nestin. Scale bars, 100 μm. c , Immunofluorescence images of iPSNs from all three cell lines were positive for neuronal marker, Tuj1. Scale bars, 100 μm. d and e , Detection and quantification of LRP1 protein levels in WT and LRP1 -KO iPSNs via Western blotting. f and g , Endocytosis of human tau in WT and LRP1 -KO iPSNs measured by flow cytometry (100 nM, 3 h of treatment). Experiments in ( f and g ) were performed in technical duplicates or triplicates over three independent experiments. All data are expressed as mean ± s.d. with individual data points shown. Data were analyzed by One-way ANOVA with Tukey’s multiple comparisons test. NS, not significant; *** P < 0.001

Article Snippet: To differentiate the cells into neural progenitor cells (NPCs), the medium was replaced to neural progenitor cell medium (Stemcell Technologies, cat# 05834) and cultured for additional 10–14 days.

Techniques: Biomarker Discovery, Derivative Assay, Gene Knockout, CRISPR, Labeling, Flow Cytometry, Immunofluorescence, Marker, Western Blot

Journal: eBioMedicine

Article Title: Global and single-cell proteomics view of the co-evolution between neural progenitors and breast cancer cells in a co-culture model

doi: 10.1016/j.ebiom.2024.105325

Figure Lengend Snippet: Neural progenitor cell differentiation. a Brightfield images displaying the morphological changes in NPCs following differentiation into midbrain dopaminergic neurons, immature neurons (after two weeks of differentiation) and maturing neurons (after five weeks of differentiation). Scale bar – 200 μm. b Sp8 confocal images showing presence of DCX, NFL, SOX1, (low) TUBB3, Nestin, KI67 in the hESC-derived NPCs. With staining showing negative expression of INA, NFH and AE1/AE3. Scale bar – 42 μm. Presence of tyrosine hydroxylase (TH) in green. Expression marked by white arrows. c Fold change of neural markers comparing midbrain neurons (n = 2 samples), immature neurons (n = 2), and maturing neurons (n = 2 samples at two weeks of maturation and n = 2, 5 weeks) to NPC protein expression SEM bar (∗p ≤ 0.05, significance was determined by unpaired t-test).

Article Snippet: Differentiation of Neural Progenitor Cells derived from XCL-1 DCXp-GFP were conducted in accordance with “Protocols for Neural Progenitor Cell Expansion and Dopaminergic Neuron Differentiation” from ATCC.

Techniques: Cell Differentiation, Derivative Assay, Staining, Expressing