Temporal expression pattern of nos2a and nos2b during zebrafish development. Analysis was performed by reverse transcriptase-polymerase chain reaction (RT-PCR) using embryos and larvae at indicated stages (hpf: hours post-fertilization). β-actin was used as a housekeeper gene control.

Spatial expression pattern of nos2a analyzed by WISH on 48, 72 and 96 hpf zebrafish. Specificity of the nos2a antisense probe (B colunn) was verified by using nos2a sense probe (A colunn).

Expression of nos2a and nos2b genes in different zebrafish adult tissues. Total RNA was extracted from tissues isolated from 6 month old zebrafish for RT-PCR using nos2a and nos2b specific primers. Primers for β-actin gene were used as positive control.

Induction of nos2a and nos2b gene expression. ZFL (A) or ZF4 (B) cell lines remained unstimulated (c) or were stimulated with LPS or PolyI:C for 8 or 20 h. Adult zebrafish (C) were challenged with LPS (5 μg in 50 μL) or PBS (as a negative control). Total RNA from cells or site of injection was reversed-transcribed and amplified to detect nos2a, nos2b, ifn1 and the β-actin control.

Induction of nos2a and nos2b gene expression after tail transection. WISH experiments are performed with nos2a (A, B) and nos2b (C, D) antisense riboprobes on 4 dpf control larvae (A, C) or on larvae on which tail transection was performed 10 h previously (B, D).