(A) Amino acid sequence alignment of human (NM_024080), mouse (NM_134252),
and rat TRPM8 (NM_134371). The alignment and text graphics were generated
using Clustal Omega software (EMBL-EBI, Hinxton, UK; https://www.ebi.ac.uk/Tools/msa/clustalo/). (B) Calcium flux
elicited by CFA (2.3 mg/ml per 180
µg/cm2) and menthol (500
µM) on GCaMP6-overexpressing cells transiently
transfected with either wild-type (WT) human (menthol n =
35, CFA n = 32) or mouse (menthol n = 6,
CFA n = 6) or the mouse-to-human mutants S921G (menthol,
n = 14; CFA, n = 16), S927A (menthol,
n = 17; CFA, n = 11), and S932T
(menthol, n = 14; CFA, n = 10). Data are
represented as the mean ± S.E.M. Statistically significant responses
using two-way analysis of variance and post-testing with the
Dunnett’s multiple comparisons test relative to mouse TrpM8-WT are
represented as *P < 0.05. mM8WT, mouse TRPM8
wild type; mS921G, mouse S921G; mS927A, mouse S927A; mS932T, mouse
S932T.