h-CaD and ERK 1/2 phosphorylation during myometrial contractions in vitro.
(A) Myometrial strips were snap frozen at specific stages during the development of contractions. The tissue was then pulverised and subjected to protein extraction. Protein (20 µg) was separated by 1D PAGE and transferred to a nitrocellulose membrane. (B) Membranes were probed with antibodies against phospho-h-CaD (1∶2000), total h-CaD (1∶2000), phospho-ERK 1/2 (1∶2000) and total ERK 1/2 (1∶2000), as well as α-SMA (1∶10000), representative images shown of 5 replicates. (C) Non-parametric analyses of the non-normalised raw data revealed a statistically significant 2-fold increase in h-CaD phosphorylation during the transition from point 1 to point 2. No significant difference was observed between contraction points 1 and 3 or 2 and 3. *p<0.05. Mr = relative molecular mass×1000.