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. 2022 Jul 24;146(2):475–491. doi: 10.1093/brain/awac273

Figure 3.

Figure 3

Sigma-1R expression is necessary for the reduction of TRPA1 whole-cell currents by S1RA. (A) Representative time course of whole-cell currents, from the HEK293 hTRPA1-tGFP stable cell line, in response to the indicated repeated voltage ramp protocol (red) and AITC (50 µM), HP = −60 mV, in CS and after S1RA pre-incubation (100 µM, 4 h). Note the slow development of current and the reduced peak amplitude after S1RA incubation. (B) Average IV relationships from HEK293-hTRPA1-tGFP cells using the protocol shown in A recorded in control (n = 10) and S1RA at different concentrations: 10 µM (n = 10), 30 µM (n = 10) and 100 µM (n = 10). Average of maximal response to AITC in each trace. (C) Dose-response curve of S1RA effects on HEK293 hTRPA1-tGFP cells in response to AITC (50 µM). Normalized current density at −60 mV with respect to CS. ***P < 0.001 (ANOVA in combination with Dunnett’s post hoc test). (D) Average IV relationships from WT HEK293 cells transiently transfected with hTRPA1-tGFP in control (n = 15), S1RA pre-incubation (100 µM, 4 h, n = 12) and in Sigma-1R silenced cells transiently transfected with hTRPA1-tGFP (S1R-KO) recorded after pre-incubation in CS for 4 h (n = 12) or S1RA (100 µM, 4 h, n = 18). Average of maximal response to AITC in each case. (E) Summary of individual and mean ± SE current density at −60 and +60 mV for the different experimental conditions shown in D. **P < 0.01 (ANOVA in combination with Bonferroni’s post hoc test). Results from transient transfections were obtained in seven independent experiments.