Fig. 1

A Real time PCR for nos1, nos2a, and nos2b in tailfin at 3, 5, and 10 dpa. B Western blot (WB) analysis of Nos2 in tailfins at 3, 5, and 10 dpa and semi-quantitative analysis of bands. C WB analysis of Nos2 in nucleus and cytoplasm of tailfins at 3, 5, and 10 dpa. Semi-quantitative analysis of bands shows nuclear to cytoplasmic protein ratio. D WB analysis of S-nitrosylated nuclear proteins in the regenerating tailfin. S-nitrosothiols were specifically labeled with TMT. An anti-TMT antibody was used to detect S-nitrosylated proteins. Neg and Pos are respectively the negative (without ascorbic acid) and positive (with S-nitrosoglutathione) controls. E, F WB analysis of S-nitrosylated nuclear proteins in tailfin following treatment with L-NAME 10 or 50 mM, 2-Phenyl-4,4,5,5-tetramethyl imidazoline-1-oxyl 3-oxide (PTIO) 3 or 10 mM, S-Nitroso-N-acetyl-dl-penicillamine (SNAP) 10 or 30 mM, or PBS (control). The dot plot shows changes in tailfin regeneration rate, here shown at 7 dpa, following drug treatments compared to control. β-tubulin was used as loading control for total or cytoplasmic proteins. Histone H3 was used as loading control for nuclear proteins. Dpa days post-amputation. N = 3 biological replicates, one way ANOVA test followed by Bonferroni’s multiple comparisons test was used to compare the means, p values shown are vs. uninjured or control, all other comparisons are not significant. Data are presented as mean values +/− SEM.