Fig. 1

Subcellular localization of PRMT5 and histone H4R3me2s during oligodendrocyte differentiation. a Prmt5 transcripts in proliferating (+GF), arrested OPC (−GF), OliNeu, or glioma cells. Scatter plots represent the average of three biological replicates, each in triplicate and normalized by the average of three housekeeping genes (18s, Wdr33, and Pja2). Values are referred as relative to those measured in OPC + GF (one-way ANOVA with Bonferroni’s multiple comparison test, *p < 0.05). b Confocal images of cells stained for PRMT5 (green), DAPI (blue), and either KI67 (red) and PDGFRα (white) in proliferating OPCs, or CNP (red) and MBP (white) in differentiating OPCs. Scale bar: 10 μm. c Relative quantification of cytoplasmic or nuclear PRMT5 in 150 PDGFRα⁺ or CNP⁺ cells. Values represent means ± sem from four biological replicates (one-way ANOVA, ***p < 0.001). d Prmt5 transcripts in three biological RNA preparations from mouse spinal cord at indicated time points, normalized as described in a and referred to levels detected at P1 (one-way ANOVA with Bonferroni’s multiple comparison test, **p < 0.01, ***p < 0.001). e Confocal images of P7 brains stained for PRMT5 (green), OLIG2 (red), MBP (white), and DAPI (blue). Scale bar: 10 μM. f Quantification of nuclear PRMT5 intensity in OPCs. Scatter plots representing the average pixel intensity of 600 (for OPC + GF and OPC − GF12 h) and 569 (OPC − GF 48 h) nuclei quantified per condition (technical triplicates of four different biological preparations). One-way ANOVA, ***p < 0.001. g Confocal image of the PRMT5-specific mark H4R3me2s (green) in PDGFRα⁺ (white) or MBP⁺ (white) cells. DAPI (blue) as nuclear counterstaining. Scale bar: 5 μm. h Scatter plots representing the average pixel intensity of H4R3me2s-stained nuclei. Quantification of 468 nuclei of cells in proliferating conditions, 543 nuclei in growth-arrested, and 493 in differentiation conditions, each from four different preparations. One-way ANOVA, ***p < 0.001. i Representative confocal image of P4 mouse brain stained for H4Rme2s (green), OLIG2 (red), and DAPI (blue). Scale bar: 5 μm. j Scatter plots representing the average pixel intensity of H4R3me2s-stained nuclei of OLIG2⁺ at the indicated time points. Two hundred nuclei were quantified at each time point (50 cells/animal and four mice per time point. One-way ANOVA, ***p < 0.001