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Fig. 5 | Acta Neuropathologica Communications

Fig. 5

From: Human induced pluripotent stem cell engineering establishes a humanized mouse platform for pediatric low-grade glioma modeling

Fig. 5

Cxcl10 absence is both necessary and sufficient for LGG formation a NF1-null iNPC cell numbers decrease (direct cell count, top; 9–20% decrease) and the percent of cleaved caspase-3+ cells increases (bottom; 8.3–20.5-fold) following ectopic Cxcl10 expression or incubation with 25 or 100 pg/mL of recombinant Cxcl10 protein. b Representative immunocytochemistry and quantification, demonstrating that ectopic Cxcl10 expression or treatment with increasing concentrations of recombinant Cxcl10 peptide induce an increase in GFAP+ astrocytic differentiation (Cxcl10: 8.3-fold, 25 pg/mL Cxcl10: 8.2-fold and 100 pg/mL Cxcl10: 20.5-fold increase), while 95–100% of the differentiated GFAP+ cells are undergoing apoptosis (cleaved caspase-3+) and 83.3–88.8% of the total number of cells undergoing apoptosis (cleaved caspase-3+) are GFAP+. c Top panel: Ectopic expression of murine Cxcl10 in 2041C > T−/− NF1-null iNPCs and iGRPs (GFP expression, Western blot). α-tubulin was used as an internal protein loading control. Bottom panel: Ectopic murine Cxcl10 expression inhibits LGG formation in Rag1−/− mice at 1mpi (H&E images). Scale bar, 1 mm. d Representative images of H&E, GFAP, OLIG2, synaptophysin and Ki67 staining of 2041C > T−/− NF1-null iNPC- and iGRP- derived LGGs in Cxcl10−/− mice at 1mpi. The number of LGG-bearing mice is indicated within the H&E panels. Scale bars: left H&E panel, 1 mm; other panels, 100 µm. e Bar graphs denoting the percent of Ki67+ cells in the lesions. Data are represented as means ± SEM, a and b one-way ANOVA with Dunnett’s multiple comparisons test, e two-tailed student’s t-test. Individual p values are indicated within each graph. ns not significant

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