Involvement of LMO4, HYPA, and KIAA1196 in the Smad pathway. (A) Following siRNA-mediated cellular knock-down targeting LMO4, HYPA, and KIAA1196, Q-PCR and reporter assays were used to analyze endogenous levels of human alkaline phosphatase (AP) mRNA and BMP-induced reporter (BMP-RE) activity in HepG2 cells, respectively. AP mRNA levels and BMP-RE activity were determined in untreated (white) and BMP7-treated (gray) HepG2 cells. (B) Following siRNA-mediated cellular knock-down targeting LMO4, HYPA, and KIAA1196, Q-PCR and reporter assays were used to analyze endogenous levels of PAI-1 mRNA and TGFβ-induced reporter (TGFβ-RE) activity in HepG2 cells, respectively. PAI-1 mRNA levels and TGFβ-RE activity were determined both in untreated (white) and TGFβ-treated (black) HepG2 cells. In all Q-PCR experiments, mRNA levels were normalized according to an internal GUS control. The specific luciferase activity was normalized using the pRL-TK vector. All results are mean values ±SE calculated from triplicates performed in at least two independent experiments.
