Skip to main content
. Author manuscript; available in PMC: 2010 Mar 29.
Published in final edited form as: Nat Med. 2008 Nov 30;14(12):1363–1369. doi: 10.1038/nm.1888

Figure 2.

Figure 2

Effect of LDN-193189 on BMP signaling and function. (a) Structures of dorsomorphin and LDN-193189. (b) Quantitative immunoblotting of PASMCs showing differential effects of dorsomorphin or LDN-193189 on BMP4–induced (10 ng ml−1) phosphorylation of Smad1, Smad5 and Smad8 (p-Smad1/5/8), with IC50 values of ~400 nM and ~5 nM, respectively. (c) Immunoblot of PASMCs treated with LDN-193189 showing differential inhibition of BMP4 (10 ng ml−1) or TGF-β (0.5 ng ml−1) signaling (IC50 ~ 5 nM and ≥ 1 μM, respectively). (d) Id1 promoter activity induced by transient transfection of COS cells with ALK2R206H or ALK2Q207D. Id1 promoter activity (BRE-Luc) was increased by 250- to 300-fold over control plasmid (pcDNA, n = 3 measurements, mean ± s.d., NS, no significant difference). (e) Impact of LDN-193189 on the transcriptional activities of caALK2R206H and caALK2Q207D mutants, expressed as percentage of full Id1 promoter activity (n = 3 measurements, mean ± s.d., *P < 0.05 versus untreated). (f) Impact of LDN-193189 (100 nM) on BMP4-induced (10 ng ml−1) osteoblast differentiation of C2C12 cells at various intervals before and after BMP4 treatment (n = 6 measurements, mean ± s.d., *P < 0.01 versus BMP4 treatment alone) and on cell viability (bottom panel). (g) Top, immunoblot for p-Smad1/5/8 and total Smad1 in PASMCs expressing the conditional caALK2Q207D transgene after infection with Ad.Cre or Ad.GFP showing the impact of pretreatment with LDN-193189 (100 nM). Bottom, immunoblot for phosphorylated Smad1/5/8 and total Smad1 in Ad.GFP- or Ad.Cre-infected PASMCs treated with BMP4 at varying concentrations.