Figure 3

Inhibition of sustained sphingosine responses by anti-TRPM3 antibody in HIG-82 cells. Data were generated by intracellular Ca²⁺ measurement. (a) Cells were pre-treated with 1:4000 dilution of pre-immune serum (pre-imm.) or anti-TRPM3 antiserum before observing responses to 20 μM sphingosine (SPH). (b) Summary normalised data from 8 independent experiments showing inhibitory effect of TM3E3 on the sustained response to SPH. (c) Lack of effect of TM3E3 on Ca²⁺ entry in cells store-depleted by 1 μM thapsigargin (TG) for 30 min in Ca²⁺-free extracellular solution followed by re-addition of 1.5 mM Ca (+Ca²⁺); representative of N/n = 24/3. (d-f) Investigation of the transient response to SPH. (d) Effect of 10 μM ruthenium red (+RR.) on the SPH response in HIG-82 cells; representative of N/n = 12/3. (e) Responses to 10 μM 4α-phorbol 12,13-didecanoate (4α-PDD) in HIG-82 cells in the presence (+Ca²⁺) and absence of extracellular Ca²⁺; representative of N/n = 16/2. (f) Representative of 3 independent experiments, responses to sphingosine (SPH, 20 μM) in CHO cells stably expressing TRPV4.