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Figure 3


Fig. 3. The RhoA/ROCKII pathway ultimately leading to an intact, dynamic actin cytoskeleton under tension has an important role in mechanically induced Runx2. (A) One hour incubation with LPA leading to increased amounts of active RhoA elicits a 2.6±0.8-fold increase in Runx2 expression (P≤0.01). Additionally, dynamic flow further induced Runx2 expression by 2.4±0.2-fold (P≤0.01) in LPA-treated cells, suggesting that active RhoA and oscillatory fluid flow act synergistically to enhance Runx2 upregulation. On the other hand, inhibiting ROCKII function significantly decreases Runx2 expression by 3.2±0.3-fold (P≤0.01) and abrogates flow induced Runx2 expression. (B) Multiple characteristics of the actin cytoskeleton downstream of RhoA and ROCKII activity influence flow induced Runx2 expression. Specifically, if we remove cytoskeletal tension by inhibiting myosin II activity, the cells lose the ability to upregulate Runx2 with flow. Furthermore, altering cytoskeletal polymerization with cytochalasin D or inhibiting cytoskeletal dynamics with jasplakinolide abrogates flow induced Runx2 expression. Error bars: s.e.m. (n≥6).





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