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Figure S1 Chemical structures of agonists (A), potentiators (B) and antagonists (C) used in this study.

Figure S2 Sequence and expression of human, rat and mouse TRPV3. (A) rat, human and mouse TRPV3 mRNA is detected in rat recombinant, human recombinant and m308k cells, respectively. A single band of the expected size was amplified by RT-PCR using species-specific TRPV3 primers, whereas no band was observed in HEK293 cells alone. The DNA ladder indicates size in bp. (B) TRPV3 protein is expressed in rat and human cell lines and not in untransfected HEK293 cells. TRPV3 protein was detected by Western blot using a TRPV3-specific Ab. The protein ladder indicates size in kDa. (C) Multiple sequence alignment of the human (Hs), mouse (Mm), and rat (Rn) TRPV3 protein sequences. Identical residues are highlighted in yellow. Differences between the two rat TRPV3 sequences are coloured in red. Ankyrin repeats and transmembrane regions are coloured blue and orange, respectively. The green box indicates the membrane-embedded region. (D) TRPV3 protein was detected by Western blot in HEK293 cells transiently transfected with plasmids expressing human or rat TRPV3. The rat reference sequence is indicated in brackets. The vertical line indicates splicing of the identical photo.

Figure S3 Inhibition of Ca2+ signaling in response to DPBA in human TRPV3 (A), rat TRPV3 (B) and mouse 308 keratinocytes (C) by the presence of increasing concentrations of TRPV3-selective antagonists. Data are displayed as percentage responses to DPBA alone. Each data point represents the mean response ± SEM from 4–6 separate experiments. (D) Inhibition of intracellular Ca2+ signalling by RR or FTP-THQ in the presence of DPBA and AA in m308k cells. Data are displayed as percentage responses to DPBA and AA in the absence of inhibitor. Each data point represents the mean response ± SEM from 2 separate experiments.

Figure S4 Schild plots of the mechanism of inhibition of Ca2+ responses to 50 μM DPBA and camphor in m308k cells by either (A) FTP-THQ or (B) Ruthenium Red.

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