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: Frontiers in Oncology

Article Title: ME2 Promotes Proneural–Mesenchymal Transition and Lipogenesis in Glioblastoma

doi: 10.3389/fonc.2021.715593

Figure Lengend Snippet: ME2 is positively correlated with mesenchymal features in GBM. (A) Gene expression heat map showing the expression of ME2 and PN and MES markers in different gliomas. (B) mRNA expression levels of ME2 detected by real-time PCR in SW1783, U251MG, LN229, and U87MG cells (n = 4, **P < 0.01). (C) Protein expression levels of ME2 in SW1783, U251MG, LN229, and U87MG cells detected by western blotting. (D) Expression of MES markers MET and YKL-40 and PN marker OLIG2 detected by western blotting in SW1783, U251MG, LN229, and U87MG cells.

Article Snippet: The primary antibodies included ME2 (Abcam, ab139686, 1:1000), MET (CST, 24294, 1:1000), E-cadherin (CST, 3195, 1:1000), N-cadherin (CST, 4061, 1:1000), vimentin (CST, 5741, 1:1000), YKL-40 (BioWorld, BS6564, 1:800), OLIG2 (Santa Cruz, sc-293163, 1:800), SREBP-1 (Abcam, 28481, 1:1000), ATP–citrate lyase (ACLY; CST, 13390, 1:1000), ACSS2 (CST, D19C6, 1:1000), AMPK (CST, 2532, 1:1000), phospho-AMPK (CST, 2531, 1:1000), and β-tubulin (MA5-11732, 1:2000; Thermo Fisher Scientific) antibodies.

Techniques: Gene Expression, Expressing, Real-time Polymerase Chain Reaction, Western Blot, Marker

Journal: Frontiers in Oncology

Article Title: ME2 Promotes Proneural–Mesenchymal Transition and Lipogenesis in Glioblastoma

doi: 10.3389/fonc.2021.715593

Figure Lengend Snippet: ME2 promotes PMT and inhibits the production of ROS in mitochondria. (A, B) Western blots confirming the expression of MET, E-cadherin, N-cadherin, vimentin, YKL-40, and OLIG2 (A) in SW1783 and U251MG cells transfected with vector or Flag-ME2 plasmids and (B) in LN229 and U87MG cells transfected with sh-EGFP or sh-ME2 plasmids. (C, D) Changes in ROS in mitochondria of SW1783 and U251MG cells transfected with vector or Flag-ME2 plasmids detected by fluorescent probe. Green fluorescence represents lipid components oxidized by ROS; red fluorescence represents nonoxidized lipid components. Fluorescence intensity statistics and analysis by Image J (*P < 0.05).

Article Snippet: The primary antibodies included ME2 (Abcam, ab139686, 1:1000), MET (CST, 24294, 1:1000), E-cadherin (CST, 3195, 1:1000), N-cadherin (CST, 4061, 1:1000), vimentin (CST, 5741, 1:1000), YKL-40 (BioWorld, BS6564, 1:800), OLIG2 (Santa Cruz, sc-293163, 1:800), SREBP-1 (Abcam, 28481, 1:1000), ATP–citrate lyase (ACLY; CST, 13390, 1:1000), ACSS2 (CST, D19C6, 1:1000), AMPK (CST, 2532, 1:1000), phospho-AMPK (CST, 2531, 1:1000), and β-tubulin (MA5-11732, 1:2000; Thermo Fisher Scientific) antibodies.

Techniques: Western Blot, Expressing, Transfection, Plasmid Preparation, Fluorescence

Journal: iScience

Article Title: Immune-mediated neurodegenerative trait provoked by multimodal derepression of long-interspersed nuclear element-1

doi: 10.1016/j.isci.2022.104278

Figure Lengend Snippet:

Article Snippet: The following antibodies were used; rabbit anti-L1 ORF1p (Novusbio, USA), Alexa Flour 488-conjugated rabbit anti-NeuN (Abcam), Alexa Fluor 647-conjugated Anti-Rabbit IgG (Jackson ImmunoResearch, USA) Alexa Fluor 647-conjugated rabbit anti-NeuN (Abcam)(1:200), anti-Olig2 (biorbyt)(1:50), Alexa Fluor 488-conjugated anti-Olig2 (Millipore)(1:50), Alexa Fluor 647-conjugated anti-Iba1 (Millipore, USA) (1:50), anti-IBA1 (LSBio, USA) (1:50), Alexa Fluor 647-conjugated anti-GFAP (invitrogen, USA) (1:50), Alexa Fluor 488-conjugated anti-GFAP (invitrogen, USA) (1:50), Alexa Fluor 647-conjugated anti-Rabbit IgG (Jackson ImmunoResearch, USA)(1:200).

Techniques: Staining, Fluorescence, Recombinant, Adjuvant, Magnetic Beads, Modification, Purification, cDNA Synthesis, SYBR Green Assay, Blocking Assay, Antibody Labeling, Enzyme-linked Immunosorbent Assay, Enzyme-linked Immunospot, Expressing, Plasmid Preparation, Software, Microscopy, Magnetofection

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: Science Advances

Article Title: Oriented cell divisions induce basal progenitors and regulate neural expansion across tissues and species

doi: 10.1126/sciadv.adz6827

Figure Lengend Snippet: ( A ) Representative images of adult cortical sections from dHET and dKO stained for markers of upper layer II/III neurons (BRN2), deeper layers IV/V neurons (CTIP2), and astrocytes (SOX9). Scale bar, 200 μm. ( B ) Quantification of cell type proportions in adult dHET cortex, dKO cortex, and dKO subcortical heterotopia band. Upper layer II/III, deeper layer IV/V neurons, astrocytes, interneurons, and oligodendrocytes cell types are identified using BRN2/Pou3f2, CTIP2, SOX9, GAD67, and OLIG2, respectively. ( C ) Representative images of adult retinal sections from dHET and dKO stained for markers of amacrines (PAX6), bipolars (CHX10), retinal ganglion cells (BRN3a/POU4F1), Müller cells (LHX2), cones (ARR3), rods (NRL), and horizontals (LIM1). Yellow dashed lines underlie the extra retinal layer of dKO retina, white dashed lines underlie the expanded ganglion cell layer of dKO retina. Scale bar, 100 μm. ( D ) Quantification of retinal cell type proportions in adult dHET and dKO retina (excluding the extra layer, gray and black bars) and in the extra layer (red bars). Error bars represent mean ± SEM. All comparisons between groups were done using a one-way ANOVA test followed by Tukey’s correction for multiple comparisons. (* P < 0.05, ** P < 0.01, *** P < 0.001, and **** P < 0.0001).

Article Snippet: OLIG2 , Rabbit , 1/500 , PhosphoSolutions , AB_2492193.

Techniques: Staining