Figure 10.

TAZ and miR-224 affect TGF-B and BMP signaling. (a) TAZ inhibits bone morphogenetic protein (BMP) 2-induced Id1-luc reporter activity. U2OS and MG-63 cells were transfected with vector or flag-TAZ, together with Id1-luc reporter plasmids. BMP2 treatment and luciferase assays were performed as described under 'Materials and methods' section. (b) TAZ inhibits TGF-β-induced SBE-luc reporter activity. U2OS and MG-63 cells were transfected with vector or flag-TAZ, together with SBE-luc reporter plasmid, and then treated with TGF-β for 12 h. (c and d) Stable expression of TAZ in U2OS (c) and MG-63 (d) cells decreases mRNA levels of p15, p21, PAI-1, Id1 and SMAD6. Cells were treated with or without TGF-β or BMP2 for 12 h, and total RNA was extracted for qRT-PCR analysis. (e) MiR-224 inhibits BMP2-induced Id1-luc reporter activity. U2OS and MG-63 cells were transfected with vector or pre-miR-224, together with Id1-luc reporter plasmids. BMP2 treatment and luciferase assays were done as described under 'Materials and methods' section. (f) MiR-224 inhibits TGF-β-induced SBE-luc reporter activity. U2OS and MG-63 cells were transfected with vector or pre-miR-224, together with SBE-luc reporter plasmid, and then treated with TGF-β for 12 h. (g and h) Stable expression of miR-224 in U2OS (g) and MG-63 (h) cells decreases mRNA levels of p15, p21, PAI-1, Id1 and SMAD6. Cells were treated with or without TGF-β or BMP2 for 12 h, and total RNA was extracted for qRT-PCR analysis. (i) Schematic model of the potential signaling mechanism of the TAZ–miR-224–SMAD4 axis