Figure 2.

Acvr1^G328V Hyperactivates BMP Signaling and Stimulates Glial Cell Proliferation

(A) Schematic depicting experiments in primary glial cells.

(B) Western blot of lysates from Acvr1^+/+ or Acvr1^floxG328V/+ (floxGV) glial cell, transduced with Ad-GFP or Ad-GFP-Cre, probed with the indicated antibodies.

(C) mRNA expression, measured by qPCR (left) and protein levels (right), assessed by western blot of Id1, Id2, and Id3, in Acvr1^+/+ or Acvr1^floxG328V/+ (floxGV) brainstem glial cells transduced with Ad-GFP or Ad-GFP-Cre. For qPCR, n = 3 experiments.

(D) Expression of Id1, Id2, and Id3, measured by qPCR in Acvr1^+/+ or Acvr1^floxG328V/+ brainstem glial cells transduced with Ad-GFP or Ad-GFP-Cre, and treated or not with 100 ng/mL noggin. n = 4 experiments.

(E) Percentage of 5-ethynyl-2′-deoxyuridine (EdU)-positive cells in GFP-negative and GFP-positive Acvr1^+/+ or Acvr1^floxG328V/+ glial cells transduced with Ad-GFP or Ad-GFP-Cre, and incubated with 10 μM EdU for 2 h. n = 3 experiments.

(F) Normalized E2F-Luc reporter activity in Acvr1^+/+ or Acvr1^floxG328V/+ glial cells, transduced Ad-GFP-Cre, relative to reporter activity in cells transduced with Ad-GFP. n = 4 experiments.

In all panels, mean + SEM is shown. ^∗p < 0.05, ^∗∗p < 0.01, ^∗∗∗p < 0.001; assessed by repeated-measures ANOVA (C–E) with Sidak (C and D), or Dunnett (E) multiple comparisons test, or paired t test (F). See also Figure S2.