Figure 4.

Acvr1^G328V and Pik3cA^H1047R Cooperate to Induce High-Grade Diffuse Gliomas in Mice

(A) Left: representative western blots of brainstem tissue lysates from postnatal day 7 or adult mice, probed with the indicated antibodies. Right: quantification.

(B) Expression of Id1 measured by qPCR, in glial cells carrying the indicated combination of alleles (“floxGV”: Acvr1^floxG328V), transduced with Ad-GFP or Ad-GFP-Cre, and treated with 100 ng/mL noggin. n = 3 experiments.

(C) Schematic of the Acvr1^floxG328V, Hist1h3b^K27M, Pik3ca^floxH1047R, and Olig2^Cre knockin alleles, and survival curves of mice carrying the indicated combinations of alleles.

(D) Representative H&E-stained brain tissue sections showing diffuse high-grade gliomas in three mice carrying the Acvr1^floxG328V, Pik3ca^floxH1047R, and Olig2^Cre alleles without (left), or with (middle and right) Hist1h3b^K27M. Scale bars, 2.5 mm.

(E) Representative immunohistochemistry images showing expression of the indicated proteins in tumors. Scale bars, 20 μm.

(F) Expression of selected genes, measured by qPCR, in tumors and matched normal brain tissue from control littermates.

(G) Representative H&E-stained sagittal brain tissue sections of NOD SCID gamma (NSG) mice xenografted in the brainstem (left) or midbrain (right) with cell lines derived from mouse tumors, at humane endpoint. Scale bars, 2.5 mm.

In all panels, mean + SEM is shown.^∗p < 0.05, ^∗∗p < 0.01, ^∗∗∗p < 0.001; assessed by unpaired t tests (A and F), repeated-measures ANOVA with Tukey multiple comparisons tests (B), or Gehan-Breslow-Wilcoxon test (C). See also Figure S4.