Fig. 4

Systemic Inflammation in nlrc3-like^/ Mutants in the Absence of Infection or Injury

(A and B) Neutrophil-specific marker mpo shows no brain neutrophils in wild-type siblings (arrow, A) but abundant infiltration in mutants (arrow, B). Lateral views, anterior to the left.

(C and D) Representative frames from time-lapse imaging in the intact 2.5 dpf embryos (see Movies S5 and S6) show wandering neutrophils (lyz:EGFP) in the brain of mutants (D) but none in wild-type (C), with reference to vasculature (kdrl:mCherry-CAAX). Dorsal views, anterior to the top.

(E and F) Higher magnification shows GFP channel alone (dotted-box region in C and D, respectively).

(G) Plot shows that high numbers of neutrophils infiltrate the brain in mutants (n = 15) but nearly zero in wild-type (n = 16; p = 0.0018).

(H and I) Representative images show live imaging of neutrophils at 1.5 dpf in wild-type sibling (H) and in mutant (I). (I) Mutant embryos show large numbers of neutrophils in brain (asterisk) and circulation (arrows). Lateral views, anterior to the top. ey, eye; mb, midbrain; ot, otic vesicle.

(J) Plot shows significantly higher numbers of circulating neutrophils in mutants (n = 4) compared with wild-type (n = 5; p = 0.0083).

(K) Neutrophils (lyz:EGFP as pseudocolored in red) are closely intermixed with the aberrantly coalescing macrophages (L-plastin expression only as shown in green) in the mutants, whereas wild-type neutrophils are dispersed and not clustered with macrophages.

n, number of embryos analyzed. Error bars show SEM. p < 0.01. All p values are two tailed.