Fig. 1.

Pharmacological inhibition of BMP type I receptors ALK2 and ALK3. A. mRNA levels of ALK2 and ALK3 determined by RT-qPCR and normalized to 36B4 gene expression. Data are expressed as fold increase relative to control condition (vehicle; VEH or DMSO, 1) and presented as mean ± SEM. B. Representative blots with the indicated antibodies and the corresponding quantification. Data are expressed as fold change relative to the control condition (vehicle; VEH or DMSO, 1) and presented as mean ± SEM. C – D.HAMP and ID1 mRNA levels determined by RT-qPCR and normalized to 36B4 gene expression. Data are expressed as fold increase relative to the control condition (vehicle; VEH or DMSO, 1) and presented as mean ± SEM. Experimental conditions: Huh7 treated with the inhibitors LDN-193189 (LDN) (500 nM), DMH2 (10 μM) or ML347 (150 nM) for 16 h (n ≥ 3 independent experiments). **p < 0.01, ***p < 0.005 and ****p < 0.0001, LDN vs. VEH, or DMH2 or ML347 vs. DMSO; ^#p < 0.05 and ^###p < 0.005, ML347 vs. DMH2 or LDN. E – F. Cell viability determined by crystal violet staining. Data are represented as percentage relative to the control group (vehicle; VEH or DMSO, 100%) and presented as mean ± SEM. Experimental conditions: (E) Huh7 treated with the inhibitors LDN-193189 (LDN) (500 nM), DMH2 (10 μM) or ML347 (150 nM) 1 h prior to APAP (20 mM, A20) stimulation for 16 h (n ≥ 3 independent experiments). (F) Primary mouse hepatocytes treated with the inhibitor DMH2 (5 or 10 μM) 1 h prior to APAP (10 mM, A10) stimulation for 16 h (n = 2 independent experiments performed by duplicate). *p < 0.05, **p < 0.01 and ***p < 0.005, A10 or A20 vs. C; ^#p < 0.05 and ^###p < 0.005, A10/20-DMH2 vs. A10/20-DMSO. (For interpretation of the references to color in this figure legend, the reader is referred to the Web version of this article.)