CD69 Is a TGF-β/1α,25-dihydroxyvitamin D3 Target Gene in Monocytes
Figure 7
Effects of Smad3 gene silencing on CD69 and 5-LO gene regulation by TGF-β/1α,25(OH)2D3.
(A) Smad3 mRNA copy numbers of untransduced THP-1, THP-1 transduced with shRNA against Smad 3 and THP-1 transduced with non target shRNA were determined by qPCR. Smad 3 mRNA levels are displayed relative to untransduced THP-1 cells. (B) Western Blot analysis of Smad3 protein in untransduced THP-1, THP-1 transduced with shRNA against Smad3 and THP-1 transduced with non target shRNA. Protein level is displayed relative to untransduced THP-1. (C) qPCR analysis of CD69 and 5-LO mRNA levels in untransduced THP-1, THP-1 transduced with shRNA against Smad3 and THP-1 transduced with non target shRNA with and without treatment with 1 ng/ml TGF-β and 50 nM 1α,25(OH)2D3 (TV) for 24 h, respectively. Results are presented relative to treated untransduced THP-1. For statistical analysis of (A), (B) and (C) one sample t test was used (*P<0.05; **P<0.01; ***P<0.001).