Fig. 4

The zebrafish intestinal tumor instigates local and distant inflammation. (A-H) Representative images of the sibling controls and tumor-bearing larvae carrying Tg(lyz:EGFP) transgene at 7 and 9 dpf. Bright-field (A,B,E,F) and EGFP (C,D,G,H) images are shown. Scale bar: 1 mm. (I-N) Representative images of whole-mount fluorescent immunohistochemistry for Lyz in the intestines of the sibling controls and tumor-bearing larvae at 7 dpf. Lyz (I,J) and Hoechst 33342 (K,L) images are shown. (I,J) The intestine is shown by dashed yellow lines. (M,N) In the merged images, Lyz and Hoechst 33324 signals are shown in green and blue, respectively. White arrows indicate representative neutrophils in the intestine. Scale bar: 100 µm. (O) The number of neutrophils in the intestines of the sibling controls and tumor-bearing larvae. The data harbors 6 biological replicates. Error bars represent means±s.e.m. Statistical significance was tested using Student's t-test (unpaired, one-tailed). (P,Q) Representative images of the livers of the sibling controls and tumor-bearing larvae carrying Tg(lyz:EGFP) and Tg(fabp10a:mCherry) at 7 dpf. Neutrophils and the liver are shown by green and magenta, respectively. White arrows indicate representative neutrophils in the liver. Scale bar: 100 µm. (R) The number of neutrophils in the livers of the sibling controls and tumor-bearing larvae at 7 dpf. The data harbors 12 biological replicates. Error bars represent means±s.e.m. Statistical significance was tested using Student's t-test (unpaired, one-tailed). (S) Liver size of the sibling controls and tumor-bearing larvae at 7 dpf. Liver size was measured from Tg(fabp10a:mCherry) images using ImageJ software. The data harbors 12 biological replicates. Error bars represent means±s.e.m. Statistical significance was tested using Student's t-test (unpaired, two-tailed). Data are representative of at least two independent experiments.