Arrival of CSLD6 to the cell apex depends on and occurs after actin, but CSLD6 enrichment precedes actin at the developing cell plate. (A) In moss gametophores, CLSD6 is enriched in punctate structures and in the cell plates during cell division (yellow arrowheads.) Insets from the boxed regions reveal the presence of fewer CSLD6 puncta in dividing cells. Scale bar, 20 µm. Scale bar for inset, 10 µm. Time stamps, hour:minute. Also see Video 1. (B) In moss protonemata, mScarlet-CSLD6 (magenta) accumulates at the cell apex (blue arrowheads) and at the site of cell division (yellow arrowheads.) Actin is labeled with lifeact-mEGFP (green), which also accumulates near the cell apex and at the site of cell division. Inset from the boxed region reveals that CSLD6 labels the plasma membrane, while actin is in the cytosol at the cell apex. Images are maximum projections of z-stacks from a time-lapse acquisition. Inset is from the medial plane. Scale bar, 20 µm. Scale bar for inset, 10 µm. Time stamps, hour:minute. Also see Video 2. (C) mScarlet-CSLD6 in control and LatB-treated protonemal apical cells. Cyan dotted line outlines the cells. Scale bar, 5 µm. Images are from the medial focal plane. (D) During branch formation, actin appeared (4:50) before CSLD6 accumulation (6:40). Cell expansion occurs (8:00) after actin and CSLD6 accumulation. Images are maximum projections of z-stacks from a time-lapse acquisition. Scale bar, 10 µm. Time stamps, hour:minute. Also see Video 3. (E) Kymographs generated along the yellow dashed line in C. In the kymographs, actin appears at the cell apex (yellow arrowheads) before CLSD6 (blue arrowheads) and cell expansion. Scale bars, 10 µm (horizontal) and 2 h (vertical). (F) In dividing protonemal cells, CSLD6 appears in the cell plate (5:30) earlier than actin (6:00). Images are single focal plane confocal images from a time-lapse acquisition. Scale bar, 5 µm. Time stamps, min:sec. Also see Video 4.