Journal: International Journal of Molecular Sciences

Article Title: Multisite Injections of Canine Glial-Restricted Progenitors Promote Brain Myelination and Extend the Survival of Dysmyelinated Mice

doi: 10.3390/ijms251910580

Figure Lengend Snippet: The phenotype of canine glial-restricted progenitors (cGRPs) in vitro. ( A ) Morphology of cGRPs observed during 14 days in culture. Notice bipolar morphology (arrow) and formation of cellular aggregates (symbol of a star). Abbreviations: BF, bright field; DIV, day in vitro. ( B ) Phenotypic characterization of cGRPs by immunocytochemistry. Glial progenitors/oligodendrocyte precursors were detected by expression of A2B5, NG2, PDGFRα, Olig1, Olig2, O4, and CNPase. Anti-Ki67 antibody was used as a proliferation marker and was detectable in approximately 17% of cultured cells. Anti-MBP and -GalC antibodies were used for the detection of oligodendrocytes. Nuclei were counterstained with DAPI. Scale bar: 100 μm.

Article Snippet: 4 °C: anti-Olig1 (1:500) (AB15620, Merck KGaA, Darmstadt, Germany); anti-Olig2 (1:500) (ABN899, Merck KGaA, Darmstadt, Germany); anti-Ki67 (1:10) (PA0118, Leica Biosystems, Wetzlar, Germany); anti-PDGFR (1:200) (sc-338, Santa Cruz Biotechnology, Dallas, TX, USA); anti-A2B5 (1:200) (MAB312R, Merck KGaA, Darmstadt, Germany); anti-GFAP (1:200) (Z0334, DAKO, Jena, Germany); anti-NG2 (1:200) (AB5320, Merck KGaA, Darmstadt, Germany); anti-CNPase (1:200) (AB9342, Merck KGaA, Darmstadt, Germany); anti-O4 (1:200) (MAB345, Merck KGaA, Darmstadt, Germany); anti-GalC (1:200) (MAB342, Merck KGaA, Darmstadt, Germany); anti-MBP (1:200) (MAB382, Merck KGaA, Darmstadt, Germany).

Techniques: In Vitro, Immunocytochemistry, Expressing, Marker, Cell Culture

Journal: International Journal of Molecular Sciences

Article Title: The Differentiation of Rat Oligodendroglial Cells Is Highly Influenced by the Oxygen Tension: In Vitro Model Mimicking Physiologically Normoxic Conditions

doi: 10.3390/ijms19020331

Figure Lengend Snippet: Quantification of immunolabeled oligodendrocyte progenitors in different cell culture conditions. Cell cultures were fixed either after 2 or 5 days in vitro. Blue bars show cells maintained in physiological normoxia (5% O 2 ) and green bars represent cells kept at the standard oxygen level (21% O 2 ). ( A ) Expression of the lineage-specific transcription factor Olig1 ( green ), which is detected in the cell cytoplasm on the 2nd DIV (white arrows) and gradually decreases during cell maturation; ( B ) Expression of the transcription factor Olig2 ( green ), which is relevant for the early stages of oligodendrocyte differentiation (white arrows); ( C ) Progenitor cells recognized by the presence on their surface of the NG2 marker; ( D ) Proliferating cells characterized by the presence of the Ki67 marker in the cell nuclei. The cell nuclei were visualized with Hoechst 33258 solution ( blue ). The scale bar is equivalent to 20 µm. The calculated differences were marked as the significant if: * p < 0.05, ** p < 0.01; *** p < 0.001, **** p < 0.0001.

Article Snippet: The list of primary antibodies used for the immunolabeling procedure included those recognizing the proliferating cells, i.e., anti-Ki67 protein (Novocastra-Leica Biosystems, Nussloch, Germany, 1:100) and anti-BrdU (Santa Cruz Biotechnology, Dallas, TX, USA, 1:100), as well as those recognizing specific markers of oligodendrocyte maturation, i.e., NG2 (Chemicon, Tokyo, Japan, 1:100), Olig1 (Merck, Kenilworth, NJ, USA, 1:1000), Olig2 (Merck, 1:500), CNP (Chemicon, 1:100), GalC (Chemicon, 1:200, Triton not used), MBP (Merck, 1:100), and PLP (Chemicon, 1:200).

Techniques: Immunolabeling, Cell Culture, In Vitro, Expressing, Marker, IF-P

Journal: Cell stem cell

Article Title: OLIG2 Drives Abnormal Neurodevelopmental Phenotypes in Human iPSC-Based Organoid and Chimeric Mouse Models of Down Syndrome

doi: 10.1016/j.stem.2019.04.014

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: Rabbit anti-OLIG1 , Phosphosolutions , Cat# 1537; RRID: AB_2492192.

Techniques: Virus, shRNA, Recombinant, Gene Expression, Fractionation, Luciferase, Software

Journal: eNeuro

Article Title: A Functionally Defined In Vivo Astrocyte Population Identified by c-Fos Activation in a Mouse Model of Multiple Sclerosis Modulated by S1P Signaling: Immediate-Early Astrocytes ( ieAstrocytes )

doi: 10.1523/ENEURO.0239-18.2018

Figure Lengend Snippet: Immunolabeling identifies astrocytes as major c-Fos-activated cells in EAE SC. Immunolabeling for GFAP, NeuN, Olig1,2,3, Iba1, CD3, and GFP on SC sections, as indicated. Percentage overlap as indicated. N.D., not detected. n = 3 animals. Scale bar, 100 μm (GFAP, NeuN, and Olig1,2,3) and 50 μm (Iba-1 and CD3).

Article Snippet: Sections were washed three times in TBS with 0.3 M glycine, permeabilized with 0.1% Triton X-100 in TBS, blocked with species-appropriate serum, and immunolabeled with chicken anti-GFAP (1:1000 dilution, Neuromics, Cat #CH22102, RRID: AB_10014322 ), rabbit anti-NeuN (1:400 dilution, Millipore, Cat #MABN140, Clone #27-4, RRID: AB_2571567 ), mouse anti-Olig1,2,3 (1:100 dilution, Neuromics, Cat #MO15059, Clone #257224, RRID: AB_1620409 ), rabbit anti-Iba1 (1:500 dilution, Wako, Cat #019-19741, Clone #NCNP24, RRID: AB_839504 ), and hamster anti-CD3e (1:500 dilution, BD Biosciences, Cat# 553057, Clone #145-2C11, RRID: AB_394590 ).

Techniques: Immunolabeling