Fig. 6

Nestin phosphorylation at Thr-315 orchestrates redistribution of cortactin and Pfn-1 to the membrane. Cells expressing WT or T315A nestin were treated with 10^–4 M ACh for 5 min, or left untreated (UT). Cellular localization of cortactin and Pfn-1 was evaluated by immunfluorescence microscopy. A Representative micrographs illustrating the effects of T315A nestin on the spatial localization of cortactin and Pfn-1 in HASM cells. The arrows indicate a single line scan to quantify the fluorescent signals in cells. The line scan graphs show relative fluorescent intensity. B Image analysis for protein localization was described in the section of Materials and Methods. Protein distribution in cells is expressed as ratio of the intensity at the cell periphery to cell interior. Data are means ± SE (n = 6 batches of culture from 3 donors). *p < 0.05; ** p < 0.01