Fig 5

(A) A confocal image of a LysC:DsRed⁺ (red) cell and a macrophage (pale green with dotted line outline) interacting with a V12RAS⁺ cell (green) (Video S8). (B, C) Images showing LysC:DsRed⁺ leukocytes establishing tethers with V12RAS⁺ cells, as observed in Tg(Fli:GFP; LysC:DsRed; kita:GalTA4; UAS:V12RASeGFP) larvae: (B) shows a tether entirely composed of V12RAS cytoplasm (green) (Video S9); (C) shows a chimeric tether composed of both V12RAS (green) and LysC:DsRed⁺ (red) cytoplasm. (D) A single image from a 3D movie (Video S10), of a neutrophil as it glides over and engulfs pieces of a V12RAS⁺ cell; (E) 3D reconstruction of a V12RAS⁺ cell clump (green cells) showing two macrophages (L-plastin⁺, magenta staining–yellow arrowheads) as they deform to engulf individual V12RAS⁺ cells. Small white arrows and dotted outline indicates lamellipodial protrusions extending over another pair of transformed cells. (F) A single focal plane (corresponding to that indicated in [E]) of the same pair of macrophages, showing V12RAS⁺ cell shaped phagosomes (asterisks), within them. One macrophage has partially enveloped another V12RAS⁺ cell with a thin lamellipodial extension (indicated by small white arrows). (G) 3D reconstruction of an anti-L-plastin stained mitfa:V12RAS-mitfa:mCherry injected Tg(BACMPO:eGFP) ⁱ¹¹⁴ larval flank region, showing the presence of mCherry⁺ (red) debris within L-plastin⁺MPO^- cells. (H) Image as in (G) but with single channel to highlight the mCherry⁺ V12RAS⁺ debris; green arrows indicate co-localization within L-plastin⁺ MPO⁺ neutrophils; blue arrowheads indicate co-localization within L-plastin⁺ MPO^- macrophages. Note that the larger clumps of debris all reside within macrophages. The grey tonal bars/domains at the bottom of (D) and (E) are the “floor” effect offered by Velocity software when generating a 3D reconstruction from a “Z-stack” of optical sections. Scale bars = 8 μm.