Fig. 1

DOT1L activity during spinal cord neurogenesis supports progenitor maintenance and differentiation of dorsal interneurons. (a) qRTPCR analysis of Dot1l in wild type lumbar spinal cords at different embryonic stages (E9.5-E13.5) normalized to E9.5 (E9.5-E11.5 n = 3 from pooling of 3 individual embryos each, E12.5 and E13.5 n = 8 from individual embryos). qRTPCR represented with mean ± SEM. P-values were calculated with unpaired, two-tailed Student’s t-test: * p < 0.05 (b) Immunostaining H3K79me2 mark (red) and DAPI (gray) in E12.5 lumbar spinal cords in control and conditional Dot1l knock-out (Dot1l-cKO) embryos. Hemicord profile highlighted by dotted white line. Scale bar: 100 μm. (c) Volcano plot of differentially expressed genes (DEG) in Dot1l-cKO lumbar spinal cord compared to control littermates (n = 5) at E12.5. Colored, genes with increased (red) and decreased (blue) expression for adjusted p < 0.05. (d) GO term analysis for biological processes of DEG in Dot1l-cKO. Scales of gene ratio and adjusted p-value reported to the bottom left side. Genes with upregulated biological processes in left column, with downregulated biological processes upon Dot1l-cKO in right column. Number of genes per term on the Y axis of the graph, total numbers up- or down-regulated on X axis. Threshold for enrichment analysis: adjusted p < 0.1. (e, left panel) Heatmap for DEGs in mutant littermates intersected with a gene list for expressed progenitor domain-specific genes extrapolated from [68]. Color-coding based on TPM z-score, scale at the bottom top left side. (e, right panel) Annotation of progenitor domain-specificity relative to the genes intersected in the heatmap [68]. (f) Scatterplot for DEGs with batch corrected TPM for interneuron markers (Olig3, Lhx2, Lhx9, Evx1, Foxd3, Isl1, Isl2, Lmx1b) in control (blue) and mutant (orange) samples, each dot corresponding to a single n in the transcriptome analysis. Error bars represent SD in the transcriptome. Adjusted p-values from the DEG are reported as * adjusted p < 0.05, ** adjusted p < 0.01