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Fig. 8. TACE mediates stretch activation of p38. (A) Stretch activates TACE. C2C12 myoblasts were stretched for 30 minutes and lysed in RIPA buffer. The cleavage rate of a peptide that contained the TACE cleavage site found in pro-TNF- by 5 µg of the cell lysate was determined as described in Materials and Methods. Data from three independent experiments were analyzed by paired t-test, and ^* indicates a difference at P<0.01. (B) Stretch activation of p38 is dependent on TACE activity. C2C12 myoblasts were pre-incubated for 30 minutes and stretched for 2 hours in growth medium that contained TAPI (5 µM) or an equal volume of DMSO (1 µl/ml) used as vehicle for TAPI. Myoblasts were then processed for western blot analysis of p38 activation. (C) Stretch activation of p38 is dependent on TACE expression. C2C12 myoblasts were transfected with a TACE siRNA duplex or a scrambled siRNA duplex as control. After 40 hours incubation, myoblasts were stretched for 2 hours, and p38 activation and TACE expression were evaluated by western blot analysis. Both the pro-TACE (upper band, 110 kDa) and processed TACE (lower band, 90 kDa) are shown. Results from three independent experiments were expressed in arbitrary unit and analyzed using a paired t-test comparing stretched samples with non-stretched samples in each group (control, scrambled siRNA and TACE siRNA). ^*Significant difference (P<0.01).

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