Journal: Cell reports

Article Title: CD44 correlates with longevity and enhances basal ATF6 activity and ER stress resistance

doi: 10.1016/j.celrep.2023.113130

Figure Lengend Snippet: (A) Bright-field microscopy images of NMR and mouse OPCs. Images were obtained 14 days after isolating OPCs (passage 0). Scale bars, 10 μm. (B) TPMs of widely used marker genes of OPC, oligodendrocyte, astrocyte, microglia, and neuron in NMR whole brain and in NMR brain-derived A2B5 + cells cultured for 19 days in OPC-A medium. (C) MDS analysis of RNA-seq data obtained from OPCs and brain of NMR, human, mouse, and rat. Eigenvalues are provided in parentheses. (D) The top heatmap shows the relative expression levels of genes that were differentially expressed in NMR OPCs compared with OPCs of all other species. The bottom heatmap shows the relative expression levels of genes that were differentially expressed in both NMR and human OPCs compared with mouse and rat OPCs (FC >3-fold and adjusted p value <0.05 by one-by-one comparison). Results of functional enrichment analyses are shown on the right side of the panel. (E) The boxplots show the relative expression levels of non-ECM-related and ECM-related genes (GO: 0030198) in OLIG2 + and NeuN + cells isolated from the brain of human, chimpanzee, and rhesus macaque. *p < 0.05 (Wilcoxon test).

Article Snippet: For evaluation of A2B5 expression levels, cells were detached from dishes using Accutase, pelleted by centrifugation (200 × g, 5 min), resuspended in 3% BSA/PBS containing 10 μg/mL anti-A2B5 antibody (R&D systems; cat. MAB1416), and incubated on ice for 1 h. Cells were then washed twice with PBS and incubated in 3% BSA/PBS containing 1:100 diluted Alexa Fluor 488-tagged secondary antibody (Thermo Fisher Scientific; cat. A-11001).

Techniques: Microscopy, Marker, Derivative Assay, Cell Culture, RNA Sequencing, Expressing, Comparison, Functional Assay, Isolation

Journal: Cell reports

Article Title: CD44 correlates with longevity and enhances basal ATF6 activity and ER stress resistance

doi: 10.1016/j.celrep.2023.113130

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: For evaluation of A2B5 expression levels, cells were detached from dishes using Accutase, pelleted by centrifugation (200 × g, 5 min), resuspended in 3% BSA/PBS containing 10 μg/mL anti-A2B5 antibody (R&D systems; cat. MAB1416), and incubated on ice for 1 h. Cells were then washed twice with PBS and incubated in 3% BSA/PBS containing 1:100 diluted Alexa Fluor 488-tagged secondary antibody (Thermo Fisher Scientific; cat. A-11001).

Techniques: Recombinant, Transfection, Protease Inhibitor, CyQUANT Assay, Proliferation Assay, Expressing, Mass Spectrometry, Software

Journal: PLoS ONE

Article Title: Fluorescent Protein-Expressing Neural Progenitor Cells as a Tool for Transplantation Studies

doi: 10.1371/journal.pone.0099819

Figure Lengend Snippet: (A) Dot plot of gated living P2 NPCs followed by respective histograms of NPCs stained for the cell surface markers CD133, NCAM, CD24 and A2B5. Gray lines represent positive staining, and black lines show isotype controls. Percentages indicate the number of positive cells (M2) within each population of stained cells in comparison to negative cells (M1). Most analyzed NPCs excluded 7-AAD viability staining solution and showed different percentages of marker expression. (B) FACS dot plots showing, in percentages, the combination of various surface neural progenitor/stem cell markers as indicated above each dot plot.

Article Snippet: To characterize NPCs by flow cytometry, 5×10 5 cells/200 μl were incubated in PBS with 0.1% bovine serum albumin (BSA) and the following primary antibodies for 30 min at 4°C in the dark to prevent internalization: anti-NCAM ((MEM-188)-PE 1∶100; Abcam, Cambridge, UK), PE/Cy7 anti-mouse CD24 (1∶100; BioLegend, San Diego, USA), anti-prominin-1-APC (1∶100; Miltenyi Biotec GmbH, Germany) and biotinylated anti-A2B5 antibody (1∶200; R&D Systems GmbH, Wiesbaden-Nordenstadt, Germany).

Techniques: Staining, Comparison, Marker, Expressing