Fig. 3

Inhibition of intramacrophage Mabs growth correlates with TNF-mediated ROS production.

(A-E) WT or tnfr morphant Tg(mpeg1:mCherry-F) embryos were iv infected with ≈150 R or S Mabs expressing tdTomato (A and B) or E2 crimson (C-E). (A) Number of infected macrophages in the caudal hematopoietic tissue (CHT) at 4 hpi. Horizontal lines indicate the mean values. (B) Average proportions of macrophages containing <5, 5 to 10, or >10 bacteria in the CHT at 1 dpi (n = 20 embryos for each group). Top panel shows confocal images of each representative class of infected macrophages. Scale bar, 5 μm. (C-E) CellRox green (green) staining of ROS production in infected embryos. (C) Confocal microscopy of ROS-positive infected macrophages. Scale bars, 10 μm. (D) Average proportions ROS-producing macrophages within the CHT in 1 dpi embryos (n = 30 embryos for each group). Error bars represent the SEM. (E) Representative fluorescence microscopy of 3 dpi-granuloma. Scale bars, 10 μm. Statistical significance was determined by one-tailed unpaired Student’s t test (A) or Fisher’s exact test of a contingency table (B and D). Data are representative of two independent experiments.