Figure 2. Enhanced response to ROS induced by relief from prolyl hydroxylation mediates cold sensitivity of hTRPA1.

DMOG-pretreated hTRPA1-WT- or hTRPA1-P394A-expressing cells were exposed to cold (a–c) or 10 μM H₂O₂ (d–g) and analysed using Ca²⁺ imaging experiments. (a–c) Representative traces of cold-induced [Ca²⁺]_i responses in hTRPA1-WT-expressing cells pretreated with or without 100 μM DMOG for 2 h (a, n=7–20 cells) and hTRPA1-P394A-expressing cells (b, n=11–16 cells) with or without 10 μM mitoTEMPO pretreatment, and statistical analysis (c, n=45–75 cells). ***P<0.001 versus non-treated hTRPA1-WT (white bar); ^#P<0.05, ^##P<0.01; one-way ANOVA with Tukey's post hoc test. (d,e) Representative traces of 10 μM H₂O₂-evoked [Ca²⁺]_i responses in hTRPA1-WT-expressing cells pretreated with vehicle (control) or 100 μM DMOG for 2 h (d, n=47–55 cells), and statistical analysis (e, n=4–8). *P<0.05; unpaired t test. (f,g) Representative traces of the [Ca²⁺]_i response evoked by 10 μM H₂O₂ in hTRPA1-WT- or hTRPA1-P394A-expressing cells (f, n=43–46 cells), and statistical analysis (g, n=5–6). *P<0.05; unpaired t test. All data are expressed as mean±s.e.m. Ctrl, control.