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. 2011 Dec 8;18(5):265–279. doi: 10.1093/molehr/gar078

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© The Author 2011. Published by Oxford University Press on behalf of European Society of Human Reproduction and Embryology

This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/2.5), which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

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siRNA knockdown of ROCK1 and ROCK2 in cultured myometrial cells causes a significant decrease in MYPT1 phosphorylation at Thr853. Sub-confluent myometrial cells were transfected with 25 nM siRNA targeted for ROCK1 (R1), ROCK2 (R2), both (R1/R2) or non-targeting (scr) and grown for a further 72 h. (A) Cell lysates were subjected to immunoblotting for ROCK1, ROCK2, total and phosphorylated MYL and MYPT1, representative blots are shown. Signals from the phospho-specific antibody when used were normalized to those obtained from the associated total antibody, and data were expressed as a percentage of the non-transfected value. (B) ROCK1 levels; (C) ROCK2 levels; (D) ppMYL (Thr18/Ser19) levels; (E) pMYL (Ser19) levels; (F) pMYPT1 (Thr853) levels and (G) pMYPT1 (Thr696) levels. Bars represent mean + SEM, n = 8. Statistical significance was determined using one-way ANOVA and Dunnett's post hoc test; **P < 0.01 compared with non-transfected cells.