Figure 4

Distribution of hDlg during mitosis. MCF10A cells at different stages of mitosis were stained with antibodies directed against total hDlg (all variants, α-NAG, Panels A-F) or antibodies raised against the alternatively spliced insert I3 (Panels G-J); both antibodies have previously been extensively validated for specificity against hDlg [18, 44]. Cells were co-stained with antibodies directed against β-tubulin (Panels D and G). The image overlays in panels F, I, and J show the relative distribution of hDlg (green) and microtubules (red). These images were produced by deconvolution of contiguous Z-sections. Panel J shows a reconstruction of a cross-section of the midbody ring structure co-stained with anti-I3 (green) and anti-β tubulin (red) antibodies. This cross-section is orthogonal to the central spindle. DNA (blue) was visualized by DAPI staining. The white arrows (Panel C) indicate the position of the cell membrane. All scale bars are 10 μm.