Figure 6.
Characterization of E6201 as an ACVR1 Inhibitor
(A) Luciferase activity in lysates from C2C12 cells transfected with the BRE-Luc reporter and stimulated overnight with 25 ng/mL BMP2, 25 ng/mL BMP6, or 5 ng/mL BMP9, and the indicated concentrations of E6201. n = 3 experiments.
(B) Western blot of lysates from C2C12 cell stimulated for 45 min with 25 ng/mL BMP2, 25 ng/mL BMP6, or 5 ng/mL BMP9, with the indicated concentrations for E6201 applied 45 min before BMP ligand addition, probed with the indicated antibodies.
(C) X-ray crystal structure showing interactions of E6201 (pink) in the ATP-binding pocket of ACVR1 (brown). Bound waters are shown as blue spheres. Hydrogen bonds are indicated by green dashed lines. Parts of strands β1 and β2 are omitted for clarity.
(D) Expression of Id1, measured by qPCR, in Acvr1+/+ or Acvr1floxG328V/+ glial cells transduced with Ad-GFP or Ad-GFP-Cre, and treated where indicated with 100 ng/mL noggin and/or 1 μM E6201. n = 3 experiments.
In all panels, mean + SEM is shown. ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001; assessed by repeated-measures ANOVA with Tukey multiple comparisons tests. See also Figure S6 and Table S3.