Dual Inhibition of Activin/Nodal/TGF-β and BMP Signaling Pathways by SB431542 and Dorsomorphin Induces Neuronal Differentiation of Human Adipose Derived Stem Cells

Figure 4

Dual inhibition of activin/nodal/TGF-β and BMP signaling pathways in hADSCs activates p38, Erk1/2, PI3K, and Akt1/2/3. hADSCs were grown in BM or BM supplemented with either activin/nodal/TGF-β pathway inhibitor SB or BMP pathway inhibitor DM or both for 7 and 14 days. The cells were harvested after 7 and 14 days and total proteins were harvested using extraction reagent. Proteins were resolved using electrophoresis and were transferred onto a nitrocellulose membrane using Trans-Blot cell. The bands were visualized by incubating the membrane with specific monoclonal antibodies followed by incubation with HRP-conjugated secondary antibody and then that with ECL reagent containing the substrate and enhancer solution. Intensity of bands was quantified using ImageJ software.