FIG. 5.

Involvement of actin cytoskeleton dynamics on mESC migration. mESCs were transfected with Arp3, TOCA, PAK, N-WASP siRNA (20 nM), or Nt siRNA (20 nM) for 24 h before 8-Bromo cAMP treatment for 24 h. (A) Oris cell migration assay was performed and stained with calcein-AM (5 μM). Fluorescence in the analytical zone was quantified with a plate reader. Data represent the mean ± SE of three independent experiments with triplicate dishes. *P < 0.05 versus control. **P < 0.05 versus 8-Bromo cAMP alone. (B) Wound healing migration assay was performed and images acquired before and 24 h after 8-Bromo cAMP treatment. The right panel depicted by bars denotes mean ± SE of three experiments for each condition analyzed by Meta Morph v.7.01 software. The example shown is representative of three independent experiments. *P < 0.05 versus control. **P < 0.05 versus 8-Bromo cAMP alone. (C) Live cell imaging microscopy shows directed migration of mESCs under influence of 8-Bromo cAMP. The right graph denotes the accumulated distance of migration analyzed by the multidimensional acquisition package of Meta Morph v.7.01 software. *P < 0.05 versus control. Color images available online at www.liebertpub.com/scd