Fig. 4.

CryAB^R120G expression decreases neprilysin levels. (A) Semiquantitative RT-PCR analysis of neprilysin mRNA. All data points were taken from cycle 24, which was in the linear range of the reaction. Neprilysin RNA was significantly reduced in both the unexercised and exercised hearts. (B) Time course of neprilysin protein reduction. Shown are the Western blots prepared from the same groups used to derive the data shown in Fig. 2C. The mice were allowed access to the wheels starting at 1 month (1m). (C) Exercise conserves normal neprilysin protein levels. The representative Western blot data and quantitation show significant conservation of normal neprilysin levels in the exercised CryAB^R120G hearts. β-Actin was used as a loading control. Volume is expressed in arbitrary units ± SE. ∗, significant difference vs. NTG (P < 0.001); ⋀, significant difference vs. unexercised TG (P < 0.001); n = 6.