Fig. 5

Role of nestin phosphorylation at Thr-315 in actin polymerization, MLC phosphorylation and smooth muscle contraction. A Extracts of HASM cells treated with wild type (WT) or T315A nestin were separated by SDS PAGE and immunobotted with antibodies against nestin and GAPDH. Data are means ± SE (n = 4–5 batches of culture from 3 donors). B HASM cells expressing WT or T315A nestin were treated with 10^–4 M ACh for 5 min, or left untreated (UT). F-actin/G-actin ratios in cells were evaluated using the fractionation assay. Data are means ± SE (n = 4–5 batches of culture from 3 donors). C MLC phosphorylation at Ser-19 in cells treated with WT or T315A nestin was assessed by immunoblot analysis. Basal and ACh-induced MLC phosphorylation was similar in cells expressing WT and T315A nestin (NS, not significant). Data are means ± SE (n = 4–5 batches of culture from 3 donors). D Expression of T315A nestin reduces airway constriction of human PLCS. Data are means ± SE (n = 4–6 slices from 3 donors). *p < 0.05; ** p < 0.01. One-way ANOVA was used for statistical analysis of A Two-way ANOVA was used for statistical analysis of B and C. Student’s t-test was used for statistical analysis of D