Preferential distribution of plectin in peripheral regions. REF-52 cells were treated with nocodazole to depolymerize MTs and induce collapse of most IFs to the cell center. Electron microscopy of peripheral cell regions after gelsolin treatment and immunogold (10 nm) labeling for plectin. Short intermediate filaments at the periphery have numerous plectin sidearms.
Electron microscopy of cytoskeletons was performed as described (Svitkina et al., 1995). Briefly, cells on coverslips were lysed as for light microscopy, treated, with recombinant gelsolin NHz-terminal domain, fixed with glutaraldehyde, tannic acid and uranyl acetate, critical point dried, and coated with platinum and carbon. Image corresponds to Fig 1c in J Cell Biol. 1996 Nov;135(4):991-1007.