Fig. 5: TAZ partners with TEAD1 to regulate Sox5 expression in chondrocytes.
From: TAZ is required for chondrogenesis and skeletal development
a Co-IP assay. 293 T cells were transfected with GFP-Sox5 or flag-TAZ, respectively. Cells were lysed after 48 h, and cell lysates were incubated with flag beads. The precipitated complexes were analyzed by western blot. b Immunofluorescent staining of TAZ and Sox5 in chondrocytes. Scale bars, 25 μm. c Cell proliferation rate of chondrocytes as shown by WST-1 assay after 48 h. d Chondrocyte lysates from Col2-Cre;TAZf/f mice and controls were immunoblotted with antibodies against Sox5 and GAPDH, respectively. The corresponding quantitative analysis of Sox5 expression was performed at right. e, f Representative images of immunofluorescence staining for Sox5 in tibia sections or chondrocytes from Col2-Cre;TAZf/f mice and controls. Scale bars, 75 μm (e) and 10 μm (f). g Chondrocytes were transfected with flag-TAZ or empty vector. After 24 h, the cells were treated with 50 μg/mL cycloheximide (CHX) for different periods of time as indicated. The levels of Sox5 in cell lysates were compared by immunoblotting. h Schematic diagram of TAZ/TEAD1-binding domains in the Sox5 promoter (BS, binding site; TSS, transcription start site). i ChIP assay. Co-occupation of TAZ/TEAD1 in the Sox5 promoter after loss of TAZ. j ChIP assay. Co-occupation of TAZ/TEAD1 in the Sox5 promoter after silence of TEAD1. k ChIP assay. Co-occupation of TAZ/TEAD1 in the Sox5 promoter after overexpression of TAZ. The experiment was independently repeated for three times. Error bars were the means ± SEM of triplicates from a representative experiment. *P < 0.05, **P < 0.01.
