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

Journal: bioRxiv

Article Title: Zfp423 , a Joubert syndrome gene, is a domain-specific regulator of cell cycle progression, DNA damage response and Purkinje cell development in the cerebellar primordium

doi: 10.1101/139691

Figure Lengend Snippet: Immunofluorescence staining ( A, B ) and in situ hybridization ( C ) of cerebellar sagittal sections at E12.5 (rostral to the left). A : the number of OLIG2+ proliferating PC progenitors is decreased in the Δ9-20 and increased in the Δ28-30, with respect to the wild type control (quantification in A’ ; * p <0.05; ** p <0.01, Welch t -test). B : CORL2+ postmitotic PC precursors are strongly reduced in both mutants ( B , and quantification in the B’ ; ** p <0.01, Student’s t -test), while the CORL2/OLIG2 ratio ( B’’ ) is significantly altered only in the Δ28-30 mutants (quantification in the graph; * p <0.05, Student’s t -test). C : Lhx5 , another marker of early PC differentiation, is downregulated in both mutants, compared with wild type controls. Size bar: 40 μM ( A, B ); 200μM ( C ). All the results are plotted as the mean ± s.e.m. of biological triplicates.

Article Snippet: Rabbit antibodies included: Calbindin (1:1000, Swant); ZFP423/OAZ XL (1:1000, Santa Cruz Biotechnology); FoxP2 (1:1000, Abcam); Phospho-histone H3 (1:800, monoclonal, Millipore); Phospholipase Cß4 (PLCβ4) (1:1000, kind gift Dr. M. Watanabe, Hokkaido University, Japan); Sox2 (Abcam, 1:250); Olig2 (1:500, Millipore); Corl2 (1:2000, Aviva System Biology); γ-H2AX (1:200, Cell Signaling); 53BP1 (1:1000, Bethyl Lab).

Techniques: Immunofluorescence, Staining, In Situ Hybridization, Control, Marker