Fig. 5. Active Rho and Rho-kinase are unable to restore ppMLC^T18/S19 in detached cells. (A) The HT-1080 cells were transfected with various cDNAs as indicated using the Effectene reagents for 20 hours in the presence of serum. Cells were either collected directly in TCA for the analysis of MLC and MBS phosphorylation or detached and incubated in the presence of serum for 3 minutes before being stopped with TCA. The results for ppMLC^T18/S19 are summarized from three independent experiments and are shown as the mean±s.e.m. relative to adherent pEGFP-transfected cells. The insert shows one of the representative experiments with the same sample orders as for the chart. (B) The analysis of pMBS^T853 in the detached transfected cells in comparison with the adherent pEGFP-transfected cells. The ppMLC^T18/S19 from the same samples is also shown.