Calcitriol attenuates liver fibrosis through hepatitis C virus nonstructural protein 3-transactivated protein 1-mediated TGF β1/Smad3 and NF-κB signaling pathways

Figure 3 Promotion of nonstructural protein 3-transactivated protein 1 in hepatic fibrosis via transforming growth factor beta 1 receptor/Smad3 and NF-kB signaling pathways. A and B: The protein levels of molecules in the transforming growth factor beta 1 (TGFβ1)/Smad3 and NF-kB signaling pathways after nonstructural protein 3-transactivated protein 1 (NS3TP1) interference or overexpression in LX-2 cells were analyzed by Western blot (n = 3); C and D: Western blot analysis of the above signaling pathway molecules in LX-2 cells after Smad3-specific inhibitor or licochalcone D treatment (n = 3); E: Coimmunoprecipitation analysis between NS3TP1 and Smad3 or p65; F: Luciferase activity analysis between NS3TP1 and the TGFβ1 receptor 1 (TGFβRI) promoter (n = 3); G: Luciferase activity analysis between TGFβ1 and the NS3TP1 promoter; TGFβ1 (L: 2.5 ng/mL, M: 5 ng/mL, H: 10 ng/mL) (n = 3); H: Luciferase activity analysis between NS3TP1 and the p65 promoter (n = 3); I: Luciferase activity analysis between p65 and the NS3TP1 promoter (n = 3). The data was represented as mean ± SE. ^aP < 0.001 pcDNA3.1 + TGFβ1R promoter vs pGL4.10 + pcDNA3.1, ^bP < 0.0001 NS3TP1 + TGFβ1R promoter vs NS3TP1 + pGL4.10; ^cP < 0.001; NS3TP1 + TGFβ1R promoter vs pcDNA3.1 + TGFβ1R promoter; ^dP < 0.01 pGL4.10 + NS3TP1 promoter vs pGL4.10; ^eP < 0.001 pcDNA3.1 + p65 promoter vs pGL4.10 + pcDNA3.1, ^fP < 0.01 NS3TP1 + p65 promoter vs NS3TP1 + pGL4.10; ^gP < 0.01 pcDNA3.1 + NS3TP1 promoter vs pGL4.10 + pcDNA3.1, ^hP < 0.0001 p65 + NS3TP1 promoter vs p65 + pGL4.10; ⁱP < 0.001 p65 + NS3TP1 promoter vs pcDNA3.1 + NS3TP1 promoter. NS3TP1: Nonstructural protein 3-transactivated protein 1; TGFβ1R: Transforming growth factor beta 1 receptor; p-smad3: Phosphorylated sekelsky mothers against decapentaplegic homolog 3; SIS3: Smad3-specific inhibitor; LD: Licochalcone D; Co-IP: Coimmunoprecipitation.