Journal: PLoS ONE

Article Title: Mesenchymal Transition and PDGFRA Amplification/Mutation Are Key Distinct Oncogenic Events in Pediatric Diffuse Intrinsic Pontine Gliomas

doi: 10.1371/journal.pone.0030313

Figure Lengend Snippet: Panel A: Radial plot showing the expression of oligodendroglial markers in the two groups of DIPG, in log2 ratios related each other. The red circle represent the median expression level of the whole population of DIPG. Group 1 expresses higher levels of oligodendroglial markers than group 2 DIPG. Panel B: Morphological oligodendroglial differenciation in group 1 tumors (HES staining, ×40). Panel C: Morphological astrocytic differenciation in group 2 tumors (HES staining ×40). Panel D: Olig2 immunohistochemistry in a group 1 DIPG showing that probably not all cells in the biopsy are tumoral (×40). Panel E: Dual immunohistochemistry for Olig2 and GFAP showing that tumor cells in mitosis are GFAP negative but Olig2 positive (×100). Panel F: Overall survival of 55 DIPG according to the presence (red) or absence (blue) of oligodendroglial differenciation. Median OS was shorter in patients with oligodendroglial type of tumors (7.73 vs 12.37, p = 0.045, log rank test).

Article Snippet: Sections were then incubated with various commercial monoclonal primary antibodies against Olig2 (AF 2418, 1/150, R/D system, CA, USA), P53 (DO-1, 1/1, Ventana) and MIB-1 (1/100; Dako, Glostrup, Denmark).

Techniques: Expressing, Staining, Immunohistochemistry

Journal: PloS one

Article Title: Transplantation of specific human astrocytes promotes functional recovery after spinal cord injury.

doi: 10.1371/journal.pone.0017328

Figure Lengend Snippet: Figure 1. Generation of two distinct types of astrocytes from human glial precursors after treatment with BMP4 or CNTF. Both BMP and CNTF promote the differentiation of hGPCs into GFAP-positive astrocytes expressing S100b but with otherwise distinct morphological and antigenic phenotypes. hGDAsBMP express lower levels of GFAP and exhibit a more compact morphology. hGDAsCNTF have a more elongated morphology and expressed high levels of GFAP. hGDAsCNTF also expressed high levels of neurite-outgrowth inhibitory chondroitin sulfate proteoglycans, phosphacan and CSPG4, as well as the transcription factor Olig2 - all of which have been found to be upregulated in glial scar

Article Snippet: Western blot analysis of chondroitin sulfate proteoglycans and Olig2 was performed as previously described [57], using anti-phosphacan monoclonal (1:1000, 3F8, Developmental Studies Hybridoma Bank), anti-CSPG4/NG2 monoclonal (1:2000, Chemicon), anti-Olig2 polyclonal (1:4000, Chemicon) and anti-b-tubulin monoclonal antibody (1:1000, Santa Cruz).

Techniques: Expressing

Journal: Nature Communications

Article Title: Metformin alters mitochondria-related metabolism and enhances human oligodendrocyte function

doi: 10.1038/s41467-025-63279-4

Figure Lengend Snippet: A Schematic representation of hESC-derived monolayer oligodendroglia differentiation protocol. B Illustrative immunofluorescence image of hESC-derived oligodendroglia at day 70 of in vitro differentiation (DIV) showing PDGFRα + oligodendrocyte precursor cells (OPCs) (magenta) and MBP + oligodendrocytes (grey) co-expressing OLIG2 (cyan). Scale bar = 100 μm. C Fold change (FC) difference of OLIG2 + MBP + oligodendrocytes after treatment with metformin or clemastine compared to their respective vehicle-treated controls (ddH2O or DMSO). n = 5 for metformin and vehicle control, n = 6 for clemastine and vehicle control where n = number of differentiations with 4 technical repeats. Kolmogorov-Smirnov normality test with Dallal-Wilkinson-Lillie for p -value, two-tailed unpaired t test. Mean ± SEM. D UMAP representation of clustered oligodendroglia. E Dot plot of selected marker genes showing cluster segregation. F Integration with adult human post-mortem spinal cord oligodendroglia snRNAseq dataset . G Cosine similarity heatmap showing similarities between the hESC-derived monolayer oligodendroglia (target, labelled mono_) dataset and the adult human spinal cord oligodendroglia (source) dataset. H Integration with the second and third trimester foetal OPC snRNAseq dataset . Source data are provided in the Source Data file. Created in BioRender. Swire, M. (2025) https://BioRender.com/j633z18 .

Article Snippet: Sections underwent H 2 O 2 treatment for 10 min at RT, washed in ddH 2 O and incubated for target retrieval for 5 min at 100 o C. Slides were washed again in ddh2O, followed by 0.1% PBS-Tween and incubated with anti-Olig2 primary antibody (Atlas antibodies, HPA003254) overnight at 4 o C. Sections were washed in 0.1% PBS-Tween and post-fixed in 4% PFA at room temperature for 30 min.

Techniques: Derivative Assay, Immunofluorescence, In Vitro, Expressing, Control, Two Tailed Test, Marker

Journal: Nature Communications

Article Title: Metformin alters mitochondria-related metabolism and enhances human oligodendrocyte function

doi: 10.1038/s41467-025-63279-4

Figure Lengend Snippet: A Schematic representation of hESC-derived cortical brain organoid oligodendroglia differentiation protocol. B Illustrative immunofluorescence images of hESC-derived oligodendroglia in organoids at day 67 of in vitro differentiation (DIV), showing PDGFRα + oligodendrocyte precursor cells (OPCs) (red) co-expressing OLIG2 (cyan) and of day 112 showing MBP + oligodendrocytes (yellow) co-expressing OLIG2 (cyan). Scale bars = 100 μm or 50 μm (zoom). C MBP + area of immunofluorescence per total organoid area after treatment with metformin compared to vehicle-treated controls (ddH2O). n = 5 biological replicates. Kolmogorov-Smirnov normality test with Dallal-Wilkinson-Lillie for p -value, two-tailed unpaired t test. Mean ± SEM. D UMAP representation of clustered organoid-derived oligodendroglia integrated with scRNAseq organoid data from (Marton et al. ). E Dot plot of selected marker genes showing cluster segregation. F Integration with adult human post-mortem cortical hemisphere oligodendroglia snRNAseq dataset . G Cosine similarity heatmap showing similarities between the hESC-derived brain organoid oligodendroglia (target, labelled with co_) dataset and the adult human cortical oligodendroglia (source) dataset. Source data are provided in the Source Data file. Created in BioRender. Swire, M. (2025) https://BioRender.com/cl7w2km .

Article Snippet: Sections underwent H 2 O 2 treatment for 10 min at RT, washed in ddH 2 O and incubated for target retrieval for 5 min at 100 o C. Slides were washed again in ddh2O, followed by 0.1% PBS-Tween and incubated with anti-Olig2 primary antibody (Atlas antibodies, HPA003254) overnight at 4 o C. Sections were washed in 0.1% PBS-Tween and post-fixed in 4% PFA at room temperature for 30 min.

Techniques: Derivative Assay, Immunofluorescence, In Vitro, Expressing, Two Tailed Test, Marker

Journal: Nature Communications

Article Title: Metformin alters mitochondria-related metabolism and enhances human oligodendrocyte function

doi: 10.1038/s41467-025-63279-4

Figure Lengend Snippet: A Schematic representation of green fluorescent protein-positive (GFP + ) human embryonic stem cell (hESC)-derived PDGFRA + oligodendrocyte precursor cells (OPCs) transplantation into the corpus callosum of Rag2 -/- : Shi/Shi P2-P4 mice. B Illustrative immunofluorescence and electron micrograph (EM) images of chimeric corpus callosum at 70 days post transplantation showing HuNu + cells (cyan)(left) or MBP + oligodendrocytes (cyan) co-localising with OLIG2 (magenta)(middle) and myelinated rodent axons (right). Scale bar = 100 μm (fluorescence) or 5 μm (EM). For control untreated mice, a mean of 16.15% ± 1.88 SEM rodent axons were myelinated. n = 11 animals. C Quantification of myelinated axons showed a significant increase after metformin treatment compared to the vehicle-treated (ddH 2 O) controls. Each dot represents an animal, n = 5 animals. Kolmogorov-Smirnov normality test with Dallal-Wilkinson-Lillie for p -value, two-tailed unpaired t test. Mean ± SEM. D UMAP representation of clustered oligodendroglia. E Dot plot of selected marker genes showing cluster segregation. F Integration with adult human post-mortem oligodendroglia snRNAseq dataset (brain and spinal cord) . G Cosine similarity heatmap showing similarities between the hESC-derived chimeric oligodendroglia (target, labelled with chi_) dataset and the adult human oligodendroglia (source) dataset. Source data are provided in the Source Data file. Created in BioRender. Swire, M. (2025) https://BioRender.com/ghxthad .

Article Snippet: Sections underwent H 2 O 2 treatment for 10 min at RT, washed in ddH 2 O and incubated for target retrieval for 5 min at 100 o C. Slides were washed again in ddh2O, followed by 0.1% PBS-Tween and incubated with anti-Olig2 primary antibody (Atlas antibodies, HPA003254) overnight at 4 o C. Sections were washed in 0.1% PBS-Tween and post-fixed in 4% PFA at room temperature for 30 min.

Techniques: Derivative Assay, Transplantation Assay, Immunofluorescence, Fluorescence, Control, Two Tailed Test, Marker

Journal: Nature Communications

Article Title: Metformin alters mitochondria-related metabolism and enhances human oligodendrocyte function

doi: 10.1038/s41467-025-63279-4

Figure Lengend Snippet: A Electron microscopy (EM) of myelinated chimeric animal axons showing a significant increase of mitochondrial area after metformin treatment compared to vehicle-treated controls, quantified ( B ) as the average of myelinated axons measured per mouse or number per individual axon measured to indicate the spread. n = 5 metformin- and n = 4 vehicle-treated animals. Kolmogorov-Smirnov normality test with Dallal-Wilkinson-Lillie for p -value, two-tailed unpaired t test. Mean ± SEM. Boxes in plots visualise median and 25th and 75th percentiles, and whiskers mark range up to 1.5 * inter-quartile ranges to show potential outliers. See Supplementary Data for exact summary statistics. Scale bar = 1 μm. C EM of chimeric animal corpus callosum showing glia with increased area of mitochondria after metformin treatment compared to vehicle, quantified ( D ) as average of glia measured per mouse or number per individual glial cytoplasm area measured to indicate the spread. Kolmogorov-Smirnov normality test with Dallal-Wilkinson-Lillie for p -value, two-tailed unpaired t test. n = 6 metformin- and n = 3 vehicle-treated animals. Mean ± SEM. Boxes in plots visualise median and 25th and 75th percentiles, and whiskers mark range up to 1.5 * inter-quartile ranges to show potential outliers. See Supplementary Data for exact summary statistics. Scale bar = 1 μm. E Gene Ontology (GO) analysis of differentially expressed genes between human embryonic stem cell (hESC)-derived oligodendroglia from metformin- and vehicle-treated chimeric animals. F Volcano plot of significantly differentially expressed genes between hESC-derived oligodendroglia from metformin- and vehicle-treated chimeric animals. pval < 0.05 and logFC > 0.5. DGE analysis was performed using MAST within Seurat, a two-sided statistical test that accounts for detection rate and zero inflation typical of single-cell RNA-seq data. Latent variables to correct for batch effects were included. P-values were adjusted for multiple comparisons using the Benjamini-Hochberg false discovery rate (FDR) method. G Violin plot of the most highly expressed specific genes in hESC-derived oligodendrocytes in metformin-treated chimeric animals. H Quantification of in situ hybridisation using NDUFA11 and EIF1 RNA probes and immunofluorescence for OLIG2 on human cells in chimera tissue with and without metformin treatment. n = 3 metformin- and 3 vehicle-treated animals with 2 ROIs of high-density HuNu + nuclei analysed per animal. Composite score = (fraction of OLIG2 + cells) x (mean number of RNA probe puncta). Kolmogorov-Smirnov normality test with Dallal-Wilkinson-Lillie for p -value, two-tailed unpaired t test. Mean ± SEM. I Western blot showing TOMM20 and CHCHD2 levels, compared to housekeeping B-TUBULIN in hESC-derived oligodendroglial monocultures treated for 7 days with metformin versus vehicle, with quantification. Mean ± SEM, two-tailed paired t test. M = metformin treated, C = vehicle treated controls. n = 3 separate differentiations. J Violin plot showing expression of EIF1 in multiple sclerosis (MS) donor oligodendrocytes with and without metformin (met) treatment versus controls (CTR). Source data are provided in the Source Data file.

Article Snippet: Sections underwent H 2 O 2 treatment for 10 min at RT, washed in ddH 2 O and incubated for target retrieval for 5 min at 100 o C. Slides were washed again in ddh2O, followed by 0.1% PBS-Tween and incubated with anti-Olig2 primary antibody (Atlas antibodies, HPA003254) overnight at 4 o C. Sections were washed in 0.1% PBS-Tween and post-fixed in 4% PFA at room temperature for 30 min.

Techniques: Electron Microscopy, Two Tailed Test, Derivative Assay, RNA Sequencing, In Situ, Hybridization, Immunofluorescence, Western Blot, Expressing

Journal: eLife

Article Title: Regional heterogeneities of oligodendrocytes underlie biased Ranvier node spacing along single axons in sound localization circuit

doi: 10.7554/eLife.106415

Figure Lengend Snippet: ( A, B ) Immunostaining of MAG between E9 and E21 ( A ). Boxes with numbers at E15 are magnified ( B ). ( C ) Double immunostaining of MAG (red) and Olig2 (green), a marker of oligodendrocyte lineage cells, at E15. ( D ) Localization of MAG (green), AnkG (red), and Nav (blue) on heminode at E15. ( E ) Ranvier node is formed by restricting a gap between adjacent two heminodes during development. Note that paranodal domains, flanking the Nav-positive nodal domain, accompany fibrous AnkG clustering in heminode but not in mature node. ( F, G ) Regional difference in internodal length appeared during the period of node formation. The axons were labeled with rhodamine dextran (red) and stained with Caspr antibody (green) at E18 and P9 ( F ). Internodal length was measured as a distance between adjacent mature/immature nodes ( G ). Boxes with numbers correspond to high-magnification images of each node. E18: n=41 internodes, N=4 chicks for tract, n=38 internodes, N=11 chicks for NL; P9: n=43 internodes, N=7 chicks for tract, n=62 internodes, N=10 chicks for NL. Scale bars: 200 µm (A), 50 µm (B), 30 µm (F, upper), 20 µm (C), and 5 µm (D and F, lower). Statistical analysis: Kruskal–Wallis test and post hoc Steel–Dwass test ( G ) was used to compare different developmental stages in the same region and different regions at the same developmental stage: *p<0.05, **p<0.01. Figure 2—source data 1. Quantitative measurements with associated statistical analyses and effect size visualizations underlying .

Article Snippet: Mouse pan Nav antibody (5 μg/ml, Sigma-Aldrich), guinea pig pan Nav antibody (5 μg/ml; ), rabbit AnkG antibody (5 μg/ml, a gift from Gisèle Alcaraz, ), mouse Caspr antibody (2 μg/ml, NeuroMab), mouse MAG antibody (2 μg/ml, Merck), rabbit Olig2 antibody (2 μg/ml, a gift from Hirohide Takebayashi, ), mouse Nkx2.2 antibody (2 μg/ml, DSHB), rat BrdU antibody (10 μg/ml, Abcam), rabbit GFP antibody (2 μg/ml, MBL), rat GFP antibody (2 μg/ml, Santa Cruz Biotechnology), and rabbit RFP antibody (2 μg/ml, Rockland Immunochemicals) were used for immunohistochemistry.

Techniques: Immunostaining, Double Immunostaining, Marker, Labeling, Staining

Journal: eLife

Article Title: Regional heterogeneities of oligodendrocytes underlie biased Ranvier node spacing along single axons in sound localization circuit

doi: 10.7554/eLife.106415

Figure Lengend Snippet: ( A ) Immunostainings of Olig2, a marker for oligodendrocyte lineage cells, and Nkx2.2, a marker for oligodendrocyte precursor cells (OPCs). Yellow and red arrowheads indicate tract and NL regions, respectively. ( B, C ) Density of developing oligodendrocytes increased especially at the NL region. Developmental changes in the density of Olig2-positive ( B ) and Nkx2.2-positive ( C ) cells at each region and their ratio between the regions. E9: N=3 chicks, E12: N=4 chicks, E15: N=4 chicks, E18: N=6 chicks, E21: N=3 chicks. ( D ) Immunostaining of BrdU, a marker for proliferating cells. Yellow and red arrowheads indicate tract and NL regions, respectively. ( E ) Cell proliferation was facilitated at the NL region. Developmental changes in the density of BrdU-positive cells at each region and their ratio between the regions. N=3 chicks for each stage. ( F ) Colocalization of BrdU (green), Nkx2.2 (red), and Olig2 (blue) signals at E12. ( G ) Proliferating cells were almost exclusively OPCs. Mutual percentages between BrdU-positive cells and Nkx2.2- or Olig2-positive cells at E12 and E14. N=4 chicks. Scale bars: 100 µm ( A, D ) and 20 µm ( F ). , ( D ) and 20 µm ( F ). Statistical analysis: Two-tailed paired t -test ( B, C, E ), Wilcoxon rank sum test ( G ): *p<0.05, **p<0.01. Figure 5—source data 1. Quantitative measurements with associated statistical analyses underlying .

Article Snippet: Mouse pan Nav antibody (5 μg/ml, Sigma-Aldrich), guinea pig pan Nav antibody (5 μg/ml; ), rabbit AnkG antibody (5 μg/ml, a gift from Gisèle Alcaraz, ), mouse Caspr antibody (2 μg/ml, NeuroMab), mouse MAG antibody (2 μg/ml, Merck), rabbit Olig2 antibody (2 μg/ml, a gift from Hirohide Takebayashi, ), mouse Nkx2.2 antibody (2 μg/ml, DSHB), rat BrdU antibody (10 μg/ml, Abcam), rabbit GFP antibody (2 μg/ml, MBL), rat GFP antibody (2 μg/ml, Santa Cruz Biotechnology), and rabbit RFP antibody (2 μg/ml, Rockland Immunochemicals) were used for immunohistochemistry.

Techniques: Marker, Immunostaining, Two Tailed Test