Figures 12.

Microtubule array and mitochondria distribution in HeLa cells expressing INF2 variants. HeLa cells transfected with eGFP-tagged wild-type INF2, T161N, and G73D variants were stained with MitoTracker (red) and anti-α-tubulin (white). The outline of the transfected cells is indicated by dotted lines. (A) Subcellular distribution of mitochondria and microtubules. Mitochondria in cells expressing the T161N variant (arrowhead) were mis-distributed to the cell periphery, while those in wild-type cells were predominantly located in the perinuclear region. In cells expressing wild-type INF2, microtubules were organized in a diffuse, radiating pattern. Cells expressing T161N and G73D variants had varying degrees of aberrant alignment and had parallel bundles in particular. Bars = 20 μm. (B) The three subtypes are classified by the following criteria: Class 1: normal subtype: oriented in an array radiating from the centrosome area near the nuclei (normal subtype). Arrow indicates microtubule organizing center. Class 2: intermediate subtype: some regions of the cytoplasm are occupied by microtubules arranged in parallel, while the remaining area is filled with a normal radiating array; and Class 3: disorganized subtype: microtubules are globally aligned in parallel (bundle formation) along the long axis of the cells. The diagram shows a proportional distribution of these phenotypes in HeLa cells. Cells from three independent transfections were classified (n = 200 cells).