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. 2021 Apr 12;12:2184. doi: 10.1038/s41467-021-22225-w

Fig. 1. GBM cells invading into white matter acquire pre-oligodendrocyte fate.

Fig. 1

a representative fluorescence image of a G144 xenograft (PDX144) collected for region-specific RNA-seq. Brain regions microdissected for FACS purification of GFP+ cells are shown, nuclei are counterstained with DAPI (blue). Scale = 1 mm. b PCA plot of rlog-normalised DESeq2 expression scores for the resulting RNA-seq libraries. n = 3 xenografts. c left panel: K-means clustering of DESeq2 expression ratios (absolute log2 expression ratio > 0.58, padjust < 0.05) for tumour cells invading into the corpus callosum (CC) or striatum (ST) relative to the tumour bulk. One sided Fisher exact test with Bonferroni multiple comparisons test. B Right panel: overlap of K-means cluster with gene signatures for oligodendrocyte progenitor cell (OPC), Oligodendrocytic, Astrocytic and Neuronal lineages as well as proliferation signatures. Colours represent -log10 of enrichment adjusted p-values. d top: DESeq2 log2 expression ratios of markers of the oligodendrocyte lineage organised by differentiation stage in tumour cells from CC and ST relative to B. bottom: schematic illustration of the maturation of oligodendrocyte lineage cells. e representative immunofluorescence staining for SOX10 (red) and the myelin marker oligodendrocyte-specific protein (OSP, grey) of GFP-labelled PDX144 tumours (green). Scale = 1 mm. f quantification of percentage of SOX10+ tumour cells in grey (GM) and white matter (WM). ≥750 cells per xenograft were counted. Mean ± SEM, n = 3 xenografts p = 0.000047. Unpaired two-tailed Student’s t test. g representative EdU (turquoise) and SOX10 (red) staining and h quantification of actively proliferating SOX10+ and SOX10− GFP+ tumour cells in the corpus callosum. Scale = 500 µm. ≥1000 cells per xenograft were counted. Mean ± SEM, n = 3 xenografts. p = 0.01, unpaired two-tailed Student’s t test. i representative SOX10+ (red)/EdU− (Turquoise) immunofluorescence staining and j quantification of GFP+ tumour cells implanted in the white matter of the corpus callosum (top) or the grey matter of the upper cortex (bottom). ≥140 cells per xenograft were counted. Scale = 100 µm, Mean ± SEM, n = 3 xenografts p = 0.008. Unpaired two-tailed Student’s t test.