FIGURE 4

Prolonged pro-inflammatory response and reduction in cancer cell numbers upon consecutive local injections of E. coli or Coley’s toxins. (A) Schematic of the experimental timeline for single/multiple locally infected cancer studies showing the region (black box) to be imaged in (B–E,G–J). (B–E) Multi-channel confocal images of the flank of cancerous Tg(kita:HRASG12V-GFP) larvae showing pre-neoplastic cells (green) at 3 dpfi after single vs. multiple local injections of control media (B,D) or E. coli(C,E). (F) Graph showing number of pre-neoplastic cells following each treatment. (G–J) Multi-channel confocal images of the flank of cancerous Tg(kita:HRASG12V-GFP;mpeg1:mCherry;tnfα:GFP) showing pre-neoplastic cells (green) and tnfα-positive macrophages (yellow) (tnfα-negative macrophages are red) at 1 dpfi (G,H) or 3 dpfi (I,J) after single vs. multiple local injections of E. coli. (G’–J’) Single-channel confocal images of the same larvae showing GFP-expressing cells (pre-neoplastic cells and tnfα-positive cells). (K) Graph showing the percentage of tnfα-positive macrophages at two timepoints for all of the injection regimes. Scale bars = 100 μm.