Fig. 8. Migration of torsinA^+/+ and torsinA^-/- MEFs. (A) A microfabrication chamber was designed for wound-healing assays to provide a precise physical reference of the original position of the wound edge during long-term observations. (a) A micropatterned coverslip was partially covered with a thin polydimethylsiloxane (PDMS) membrane. (b) A thin layer of chrome was deposited over the entire surface of the coverslip and membrane. (c) The coverslip was placed in a Petri dish and covered with a suspension of cells. (d) The cells were allowed to form a confluent monolayer on the glass and the thin PDMS membrane. (e) The wound was initiated by peeling off the PDMS membrane, which removes all cells except those on the chrome layer. (B) Confluent monolayers were incubated in serum-free medium for 48 hours, then a wound was made in the monolayer and 5% serum was added (Gomes and Gundersen, 2006). Marked regions in the wound were photographed under phase-contrast microscopy, sequentially at intervals up to 26 hours post-wounding and the migration of cells into the cleared space was monitored at different time points. Digital images were taken at 10x magnification and evaluated using MetaVuE software with Microsoft Excel. Photographs of fibroblast monolayers were taken under phase-contrast microscopy 4, 8 and 26 hours after wounding (a representative region is shown). Scale bar: 150 µm. (C) The migration of the leading edge of cells was measured (in µm) from fixed points on the wound edge. Values represent mean ± s.e.m. (error bars) of measurements in six regions per time point for each genotype from seven experiments. Differences between torsinA^+/+ and torsinA^-/- cells were significant at all time points (P<0.0001, ANOVA).