Fig. 2: Radiation-stress promotes endothelial and pericyte marker expression in glioma cells. | Nature Communications

Fig. 2: Radiation-stress promotes endothelial and pericyte marker expression in glioma cells.

From: P300 promotes tumor recurrence by regulating radiation-induced conversion of glioma stem cells to vascular-like cells

Fig. 2: Radiation-stress promotes endothelial and pericyte marker expression in glioma cells.

a Heatmap of relative expression of endothelial, pericyte, stemness and EMT genes in control (0 Gy), fractionated (2 Gy X 4) and single dose (8 Gy) radiated glioma cells. N = 3 biological replicates, * and ** indicates p < 0.05 and p < 0.005, unpaired two-tailed t-test. b Heatmap of relative expression of endothelial, pericyte and stemness markers in radiated vs control glioma cells from multiple patient-derived gliomasphere lines. c, d mCherry expression in control and 2,3, and 7-days radiated cells. Flow-cytometric quantitation of percentage of mCherry+ cells. Error bars represent mean ± SD, N = 3 biological replicates. ** indicates p < 0.005, one-way ANOVA. e Heatmap of relative expression of endothelial, pericyte and stemness genes in CD133 + (GSC) and CD133- (non-GSC) fractions 3- and 7-days postradiation. N = 3 biological replicates, * and ** p < 0.05, and p < 0.005, one-way ANOVA, post hoc t-test. f Immunostaining of endothelial (CD31, CD144/VE-CADHERIN) and pericyte (DES, aSMA) markers in control and radiated CD133 + and CD133- fractions. Scale bars, 50μm. g Quantitation of mCherry+ cells in control and radiated CD133 + and CD133- fractions. N = 3 biological replicates, error bars represent mean ± SD, ** p < 0.005, unpaired two-tailed t-test. h Immunostaining of VE-CADHERIN and DESMIN in GFP + tumor cells in control and radiated xenografts. Scale bars, 25 μm. White arrows point to GFP + marker+ cells in each image. i, j Immunostaining of mCherry and GFP in control and radiated tumor xenografts. Flow-cytometric quantitation of mCherry+ cells normalized to total number of GFP + tumor cells in each group. Error bars represent mean ± SD. N = 10 biological replicates, ** indicates p < 0.005 unpaired two-tailed t-test. Scale bars, 100μm k, l. Immunostaining of VE-CADHERIN and DESMIN in GFP + tumor cells in murine GBM model. Arrows point to marker+ GFP + cells. Scale bars, 25μm. Graphs show quantitation of number of GFP + marker+ cells in the tumor mass per section. N = 3 mice, * and *** indicates p < 0.05 and p < 0.0005 unpaired two-tailed t-test.

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