Fig 2: TRPV3 and ANO1 interaction in mouse primary skin keratinocytes.a, b Representative traces of 10 mM camphor-induced currents in primary keratinocytes isolated from WT (a) and TRPV3-/- (b) mice. All the recordings were performed with standard NMDG-Cl bath and pipette solutions (with 100 nM free calcium in the pipette). The holding potential was -60 mV and ramp-pulses were applied from -100 to +100 mV for 300 ms duration every 3 s. c Comparison of current–voltage relationships. N equals 5 for WT (black) and 4 for TRPV3-/- (gray) keratinocytes. Error bars indicate SEM. d Comparison of densities of the camphor-induced currents in WT (black) and TRPV3-/- (gray) keratinocytes at +60 and -60 mV (means ± SEM). Statistical significance was determined with a Mann–Whitney test. e A representative trace of 10 mM camphor-induced currents with Ani9 inhibition in a WT keratinocyte. A representative trace of 10 mM camphor-induced currents with Ani9 inhibition in a WT keratinocyte. The holding potential was -60 mV and ramp-pulses were applied from -100 to +100 mV for 300 ms duration every 3 s.

Fig 3: TRPV3 and ANO1 interaction in NHEKs.a, b Representative traces of camphor-induced currents in NHEKs using an NMDG-Cl bath solution containing 148 mM chloride (a) or an NMDG-aspartate bath solution containing 4 mM chloride (b). The pipette solution contained 140 mM NMDG-Cl and 100 nM free calcium. The holding potential was -60 mV and ramp-pulses were applied from -100 to +100 mV for 300 ms duration every 5 s. c Comparison of current–voltage relationships of the currents at arrowheads in (a, b). d Calcium imaging of NHEKs upon camphor application with and without (Ca2+ (-)) extracellular calcium. e Comparison of camphor-induced peak currents at -60 mV in an NMDG-Cl bath solution with 2 mM (Ca2+ (+)) or without (Ca2+ (-)) extracellular calcium (means ± SEM, N = 5). Statistical significance was determined with a Mann–Whitney test. f Camphor-induced current with an ANO1 inhibitor, Ani9, using an NMDG-Cl bath solution containing 148 mM chloride.

Fig 4: TRP channels and ANOs expression in NHEKs.a RT-PCR of TRPs and ANOs in NHEKs. Uncropped gel images are shown in Suppl. Fig. 11. b Western blot of ANO1 in NHEKs. The predicted band size of ANO1 is 114 kDa. c, d Calcium imaging in NHEKs. Cam Camphor, a TRPV3 agonist; GSK GSK1016790A, a TRPV4 agonist; Ca2+ (-) Calcium-free bath solution. e Comparison of current–voltage relationships. The red line indicates current–voltage relationship of basal current in NHEKs using a standard bath and pipette solution. The blue line indicates current–voltage relationship in TRPV6-expressing HEK293T cells using a standard bath solution and NMDG-Cl pipette solution. The holding potential was -60 mV and ramp-pulses were applied from -100 to +100 mV for 300 ms duration every 5 s.