Fig. 4
Plectin deficiency affects the distribution of actin filaments (AFs) and vimentin filaments (VFs) in immortalized mouse astrocytes. A Immunolabeled AFs (Ai,ii) and VFs (Aiii,iv) in Plec+/+p53−/− (left panels) and Plec−/−p53−/− (right panels) astrocytes. Boxed areas in whole-cell images are shown enlarged below the corresponding micrographs. Cell outlines are depicted in white, the border of the subplasmalemmal (SPL) area (the area between the cell outline and the line calculated by reducing the cell area by 20%) is demarcated by green puncta. Cell nuclei and perinuclear (PN) region (2 × the size of the nucleus area) are outlined in yellow by a line and puncta, respectively. Scale bars: 10 µm. B Statistical analysis of cytoskeletal filament distribution. Note that compared with Plec+/+p53−/− astrocytes, in Plec−/−p53−/− astrocytes, AFs are retracted from the SPL area and are more parallel and more bundled throughout the cell cytoplasm (Bi–iii), whereas VFs are more abundant in the SPL area, less abundant in the PN area, less parallel and more bundled in the cytoplasm (Biv–vi). **P < 0.01, ***P ≤ 0.001; Mann–Whitney U test. The data were obtained from astrocytes isolated from two mice per genotype. Experiments were performed in duplicate. n indicates the number of cells analyzed