Smad7 is inactivated through a direct physical interaction with the LIM protein Hic-5/ARA55.
After blocking with 5% non-fat milk, the membranes were transferred to another container and incubated with the primary antibodies of alkaline phosphatase (ALP, ab83259), Runt-related transcription factor 2 (Runx2, ab23981), Osteopontin (OPN, ab8448), E-cadherin (ab40772), N-cadherin (ab18203), Vimentin (ab16700), ZEB1 (ab124512), Snail (ab82846), Osteocalcin (Ocn, ab93876), proliferating cell nuclear antigen (PCNA, ab18197), Cyclin A (ab181591), Cyclin E1 (ab71535), cyclin-dependent kinase 2 (CDK2, ab64669), Cyclin D1 (ab134175), CDK4 (ab199728), Wnt3a (ab28472), Wnt5a (ab229200), Notch 1 (ab52627), Notch 2 (ab8926), Notch 3 (ab23426), p-Smad2 (ab53100), Smad2 (ab33875), Smad4 (ab40759),
Smad7 (ab216428), and GAPDH (ab181602) at 4[degrees]C overnight.
A phase 2 study showed that reduction of the TGF-[beta] signaling inhibitor
SMAD7 by administration of an antisense oligonucleotide (mongersen) resulted in a significant improvement in Crohn's disease (CD) activity compared with placebo [10, 11].
Ponnusamy et al., "Enhancer of zeste homolog 2 inhibition attenuates renal fibrosis by maintaining
Smad7 and phosphatase and tensin homolog expression," Journal oftheAmerican Society of Nephrology, vol.
Arellanes-Robledo et al., "Mechanisms of action of acetaldehyde in the up-regulation of the human [alpha]2(I) collagen gene in hepatic stellate cells: Key roles of Ski, SMAD3, SMAD4, and
SMAD7," The American Journal of Pathology, vol.
He et al., "Prolyl oligopeptidase attenuates hepatic stellate cell activation through induction of
Smad7 and PPAR-[gamma]," Experimental and Therapeutic Medicine, vol.
Expressions of collagen type I (COL I), collagen type III (COL III), [alpha]-smooth muscle actin (a-SMA), fibronectin (FN), TGF[beta]1, TGF[beta] type I receptor (TGF[beta]RI), TGF[beta] type II receptor (TGF[beta]RII), Smad2, Smad3, phosphorylated Smad2 (pSmad2), phosphorylated Smad3 (pSmad3), Smad4,
Smad7, GR, VDR, RXR, and GAPDH (Abcam, Cambridge, MA, USA) in the lung tissues were demonstrated using Western blot analysis and immunochemistry technique according to the manufacturer's instructions as previously described [8].
Brivanlou, "
SMAD7 directly converts human embryonic stem cells to telencephalic fate by a default mechanism," Stem Cells, vol.
Komers et al., "miR-21 promotes renal fibrosis in diabetic nephropathy by targeting PTEN and
SMAD7," Clinical Science, vol.