LZTS2 enriches at junctions proximal to the apically resolving intercellular bridge during abscission. (A) Confocal image of intercellular bridge (acetylated tubulin, cyan) flanked by LZTS2 enrichment (magenta) in α-cat-WT restored α-cat KO MDCK cells. Orthogonal (x-z) views shown below reveal that the midbody-region/abscission site localizes apically and in close proximity to LZTS2 junctional enrichment. Scale bar, 3 μm. (B) Schematic representation of A. (C) Higher magnification view of intercellular bridge shown in A (white box inset, i). Arrowheads denote midbody-region. (D) Confocal images of filter grown MDCK cells fixed and immunostained for LZTS2 (magenta) and acetylated tubulin (cyan). Images capture various (inferred) stages of cytokinesis and abscission. Note LZTS2 enriches at punctate junction structures that surround the intercellular bridge (note five-point ring structure, magenta arrows). (E) Confocal images of filter-grown MDCK α-cat KO cells expressing GFP-α-cat-WT (upper panel) or GFP-α-catΔM1 (lower panel). Images from fixed cells captured during late stages of cytokinesis. (F) Graph shows relative LZTS2 midbody enrichment, with each symbol representing the intensity ratio between midbody (Ac-Tubulin proximal, magenta arrowhead) and adjacent adherens junctions (small magenta arrows). Error bars reflect ± SD. Significance by t test where *P < 0.05. (G) Confocal images of dividing GFP-α-cat-WT (left panels) or GFP-α-catΔM1 (right panels) (maximum intensity projection; eight z-planes) immunostained for LZTS2 and acetylated tubulin to mark intercellular bridge. Scale bar, 5 μm. Asterisks (*) show cytoplasmic LZTS2 signal, which appears depleted in α-catΔM1 compared with α-cat-WT expressing cells (as in Fig. 7 A). Insets (i) (maximum intensity projection from 2 z-planes) show expanded view of intercellular bridge region, where LZTS2 puncta (white arrows) appear in close proximity to microtubules, particularly in α-cat-WT compared with α-catΔM1 cells (gray arrows).