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Cygb is required for NO formation through molecular interactions with iNOS. a Immunostaining of Cygb in mouse airway epithelium visualized by immunofluorescence. Scale bar = 10 μm. b Quantification of cilia length from wt and Cygb mutant (ko) mouse (n = 4). Means are ± SD. Student’s t test, two-tailed, **P < 0.01. c Chemiluminescence detection of NO production by triiodide assays using mouse trachea lysates as samples comparing wt to ko with injection volume indicated. On the right, quantification of nitrite concentration normalized by protein amount in each tissue (n = 4). Means are ± SD. Student’s t test, two-tailed, *P < 0.05. d Reaction of 8 µM ferric CYGB in the presence of 250 µM NADPH. Increased absorbance at around 543 and 575 nm is consistent with the reduction of CYGB and formation of the CYGB Fe2+-O2 complex. On the left, the reaction with CYGB and NADPH only. On the right, the same reaction in presence of 12.5 nM mouse iNOS. e Traces indicating the reduction of ferric CYGB in the presence of different reducing systems: zebrafish Nos2b, mouse iNOS or NADPH alone. f L-Citrulline assay indicating NO production by iNOS in presence or absence of CYGB (n = 3–5). Means are ± SD. Student’s t test, two-tailed, **P < 0.01, ****P < 0.0001. Source data are provided as source data file.
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