Fig. 2 | Nature Communications

Fig. 2

From: Atrx inactivation drives disease-defining phenotypes in glioma cells of origin through global epigenomic remodeling

Fig. 2

Atrx deficiency modulates mNPC differentiation state. a Panther analysis of over expressed genes in Atrx- mNPCs reveals GO functional associations with development, signal transduction, and cellular motility (GPCR: G protein-coupled receptor). b relative transcript levels (RT-qPCR) of Gfap (red), Tubb3 (blue), and Olig2 (green) in Atrx+ and Atrx- mNPCs (3 replicates per genotype) subjected to differentiation in vitro. Cells in proliferation conditions were sampled in suspension (Susp) and adherent to laminin (Adh), and then in differentiation conditions (Diff) after 1, 3, and 5 days (d). In all cases, data are scaled relative to the highest expression level for each marker across the sample set. Error bars reflect standard deviation. c western blot showing levels of histiogenic markers in Atrx+ and Atrx− mNPCs in proliferation media (P) and in differentiation media for 1, 3, and 5 days (d1, d3, d5; Vinculin loading control). d, e Tubb3 (green) immunofluorescence images (d) and quantification (e) showing reduced neuronal processes and a lower number of Tubb3+ cells in Atrx- mNPCs after 5 days in differentiating conditions. In upper plot, data represent the average percentage of Tubb3+ cells counted in 15 fields at ×20 magnification for each genotype; in lower plot, data represent the average length of Tubb3+ protrusions, 30 cells/genotype analyzed (DAPI counterstain; error bars reflect standard deviation; P values determined by unpaired two-tailed t-test). f heat map showing how differentially transcribed genes in Atrx- mNPCs (upregulated and downregulated) are expressed in distinct astrocytic and oligodendrocytic precursor compartments. Embryonic days 14.5, E16.5, and E18.5 (E14.5, E16.5, and E18.5) and postnatal days 4 and 7 (P4 and P7) are displayed