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. 2020 Feb 7;21(4):e49700. doi: 10.15252/embr.201949700

Figure 4. Loss of RacGAP1/MgcRacGAP/CYK4/Tum prevents mitotic rounding and causes cell extrusion and apoptosis.

Figure 4

  1. Nuclear localised Tum‐GFP becomes cytoplasmic in mitotic cells, including a distinct plasma membrane localisation (arrows). Note also localisation to the cytokinetic ring (asterisk) and remnant mid‐bodies/ring canals in interphase cells. Quantification shown on the right (n > 7 independent samples per genotype; mean ± 1 SD shown). Scale bars ˜2 μm.
  2. A stripe of expression of ptc.Gal4 UAS.tum‐RNAi in the wing disc leads to basally extruded cells that undergo apoptosis, marked by Dcp1 immunostaining, similar to UAS.pbl‐RNAi. Quantification shown on the right (n > 10 independent samples per genotype; mean ± 1 SD shown). Scale bars ˜20 μm.
  3. Phosphorylated Myosin‐II regulatory light chain (p‐MLC; red) detected by antibody staining reveals high levels around the cortex of rounded mitotic cells in wild‐type wing epithelial cells, but not those cells homozygous mutant for tum 347, marked by expression of GFP in single‐celled clones (MARCM system). pH3 staining (blue) marks mitotic cell chromosomes. Arrow indicates a GFP‐positive single‐cell tum 347 mutant clone. Quantification shown on the right (n > 3 independent samples per genotype; mean ± 1 SD shown). Scale bars ˜20 μm.