Fig. 2-S1

TRPA1b and TLR7 nonsense mutations in the zebrafish.

(A) The 7 bp deletion in the trpa1b coding sequence generated a premature stop codon at 1412 bp. (B) Amino acid sequences of the WT and mutant Trpa1b proteins. (C) In situ hybridization probing for trpa1b mRNA in WT 3dpf larval zebrafish, demonstrating expression in RB neurons. (D) Schematic of the Trpa1b protein structure, demonstrating that a truncated protein (in the unlikely event that it was translated) would lack a critical cysteine residue required for agonist binding (Macpherson et al., 2007). (E) Trpa1b^-/- nonsense mutants locomoted more in response to increasing temperatures at levels equivalent to their WT/heterozygous siblings. (F) Normal AITC behavioral responses (increased locomotion) are abolished in trpa1b^-/- nonsense mutants. (G) The 1 bp deletion in the tlr7 coding sequence generated a premature stop codon at bp 665. (H) Amino acid sequences of WT and mutant Tlr7 proteins. (I) Schematic of the Tlr7 protein, demonstrating that a truncated protein (in the unlikely event of translation) would lack critical functional domains. (J) In situ hybridization probing for tlr7 mRNA in WT 3dpf larval zebrafish. No tlr7 expression was observed in RB somatosensory neurons. Premature stops are denoted with red highlighting. EC = extracellular, IC = intracellular, TM = transmembrane domain, LRR = leucine rich repeat, TIR = Toll Interleukin-1 Resistance domain. C and N denote c- and n-termini. (E), Student’s t-test.