xcl-1 ipscs  (STEMCELL Technologies Inc)

Journal: Cell reports

Article Title: Opposing roles of p38α-mediated phosphorylation and PRMT1-mediated arginine methylation in driving TDP-43 proteinopathy

doi: 10.1016/j.celrep.2024.115205

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Article Snippet: Human male XCL-1 iPSCs , XCell Science , XCL-1 iPSC.

Techniques: Virus, Recombinant, Protease Inhibitor, Membrane, Western Blot, Stripping, Modification, Transfection, Fractionation, Cell Culture, Lysis, Magnetic Beads, Electron Microscopy, Staining, Bicinchoninic Acid Protein Assay, Kinase Assay, LDH Cytotoxicity Assay, Mutagenesis, CyQUANT Assay, Sequencing, Negative Control, DNA Sequencing, Plasmid Preparation, Software, Imaging

Journal: Cell reports

Article Title: Opposing roles of p38α-mediated phosphorylation and PRMT1-mediated arginine methylation in driving TDP-43 proteinopathy

doi: 10.1016/j.celrep.2024.115205

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Article Snippet: On Day 34 of differentiation iMNs derived from human male XCL-1 iPSCs (XCell Science) were subjected to proteotoxic stress by treatment with 0.5μM MG-132 (Millipore Sigma) plus 0.1% DMSO or 1μM VX-745 for 24 h. On day 35 cells were harvested and subjecyed to Sequential extraction of insoluble protein aggregates.

Techniques: Virus, Recombinant, Protease Inhibitor, Membrane, Western Blot, Stripping, Modification, Transfection, Fractionation, Cell Culture, Lysis, Magnetic Beads, Electron Microscopy, Staining, Bicinchoninic Acid Protein Assay, Kinase Assay, LDH Cytotoxicity Assay, Mutagenesis, CyQUANT Assay, Sequencing, Negative Control, DNA Sequencing, Plasmid Preparation, Software, Imaging

Journal: bioRxiv

Article Title: The YTHDF Proteins Shape the Brain Gene Signatures of Alzheimer’s Disease

doi: 10.1101/2024.10.23.619425

Figure Lengend Snippet: (A) Brightfield image of Cas9-expressing NPCs used for CRISPR experiments. (B) MAP2 and DAPI staining of NPCs. NPC-Cas9hy p7 was seeded as single cells in a 24-well plate pre-coated with GFR-Matrigel, at 200,000 cells per well. Cell culture medium NPC + Hygromycin (100 ug/ml) was changed every other day. Cells were fixed at d9 with 4% PFA for 15 min and stained for MAP2. (C) Gel image of T7 Endonuclease I assay result. Multiple cleavage products in the KO cells indicate genome editing at the intended site. Band intensities of full-length and cleavage products were quantified using Fiji, and gene editing efficiency was calculated using the following equation: 100 x (1 – (1-fraction cleaved)1/2) (see Methods). (D) Western blot results confirm the perturbation effects for m6A-associated RBPs. Whole-cell lysates from KO or NT control cells were subjected to western blot analysis using the Jess Simple Western platform. Digital gel images are shown for the 6 designated targets, probed for both Actin (red) and antibodies for the corresponding KO target (black). Actin-normalized intensities of KO targets are shown at the bottom.

Article Snippet: Neural progenitor cells (NPCs) derived from XCL-1 iPSCs were obtained from Stem Cell Technologies.

Techniques: Expressing, CRISPR, Staining, Cell Culture, T7EI Assay, Western Blot, Control, Simple Western