Figure 6.

Analysis of the TGFβ1 promoter region revealed TGFβ1 as a direct target gene of SRF. A, B) SRF binding to the SRE site in the TGFβ1 promoter region was demonstrated by the EMSA assay in vitro (A) and verified by the ChIP assay with sequencing assessment in vivo (B). C) Addition of SRF activated luciferase activity driven by TGFβ1 promoter, and mutation of the SRE (solid gray bar) diminished the SRF-mediated reporter activity. Data represent the average of 3 experiments with triplicate measurements in every experiment. D) A significant decrease in the SRF protein expression was achieved in the cardiac-specific SRF-knockout heart. E) In parallel with the decline in the SRF expression level, repressed TGFβ1 transcript was detected in the SRF-null heart. qPCR data represent the average of 10 mice/group.