TGF-β stimulates SMAD7 expression via FOXO3.

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TGF-β stimulates SMAD7 expression via FOXO3. TGF-β stimulates SMAD7 expression via FOXO3. (A) Real-time qRT-PCR evaluations on the NHLF cells after stimulation with rTGF-β (10 ng/ml) and rCHIT1 (250 ng/ml) with (+) and without (−) siRNA silencing of FOXO3. (B) HEK293 cells transfected with luciferase-tagged SMAD7 promoter construct with (ProSMAD7WT; WT) and without (ProSMAD7▽(FOXO3); mutant form) FOXO3 consensus sequence were stimulated with rTGF-β and/or rCHIT1, and then luciferase activity was measured. (C) Western blot evaluations on the pFOXO3 and FOXO3 expression in the lungs of WT (−), TGF-β Tg (+), CHIT1 Tg (+), and CHIT1/TGF-β Tg mice. Whole-lung lysates were further separated into cytoplasmic and nuclear fractions and subjected to the immunoblots using anti-pFOXO3 and FOXO3 antibodies. Values in panels A and B represent the mean ± SEM of triplicated evaluations in a minimum of two separate experiments. Panel C is a representative immunoblot of three separate experiments. H3, histone H3, used as internal control of nuclear protein. **P < 0.01, ***P < 0.001, one-way ANOVA. ns, not significant. Chang-Min Lee et al. LSA 2019;2:e201900350 © 2019 Lee et al.