FIGURE 4.

Enhanced proteolytic processing of TGFβ1 and TNF, and activation of Smad signaling in Timp3^−/− cultures. A, detection of mature and cleaved TGFβ1, total and phosphorylated Smad2/3, full-length and cleaved TNF, and the corresponding β-actin as the loading control, in Ang II (1 μm)-treated or PE (1 μm)-treated (24 h) co-cultures of the indicated genotypes. The dotted line indicates different exposures of the same blot. B, ELISA was performed to measure active TGFβ1 protein released into the conditioned media of neonatal cardiomyocytes, cardiofibroblasts, or co-cultures. C, in vivo levels of cleaved TGFβ1 protein measured by ELISA in tissue homogenates of WT and Timp3^−/− hearts, 6 h after aortic banding. *, p < 0.05 compared with the corresponding sham groups; ‡, p < 0.05 compared with WT-AB.