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. 2018 Apr 18;9:1529. doi: 10.1038/s41467-018-03974-7

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© The Author(s) 2018

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Fig. 6 — SUMOylation had no effect on capsaicin-, proton- and voltage-dependent activation of TRPV1 channels. a, b Representative traces for currents at −60 mV evoked by increasing concentrations of capsaicin (Cap) as indicated for CHO-K1 cells that expressed Flag-tagged rat TRPV1 (Flag-TRPV1) (a), Flag-TRPV1+GFP-SUMO1+Ubc9 (b), Flag-TRPV1-K822R (c), and Flag-TRPV1-K822R+GFP-SUMO1+Ubc9 (d). The current–voltage relationships shown below the traces are for basal (black) and 0.03 µM capsaicin-stimulated (red) conditions. e Concentration–response curves for capsaicin-evoked currents. Solid lines indicate fits with the Hill equation. Data points are means ± s.e.m., with n = 8 for TRPV1 (black); n = 6 for TRPV1+SUMO1+Ubc9 (blue); n = 6 for TRPV1-K822R (olive); n = 6 for TRPV1-K822R+SUMO1+Ubc9 (pink). f–i Representative traces for currents at −60 mV evoked by solutions with decreasing pH values as indicated for CHO-K1 cells that expressed Flag-TRPV1 (f), Flag-TRPV1+GFP-SUMO1+Ubc9 (g), Flag-TRPV1-K822R (h), and Flag-TRPV1-K822R+GFP-SUMO1+Ubc9 (i). j Concentration–response curves for proton-evoked currents. Solid lines indicate fits with the Hill equation. Data points are means ± s.e.m., with n = 7 for TRPV1 (black); n = 9 for TRPV1+SUMO1+Ubc9 (blue); n = 7 for TRPV1-K822R (olive); n = 7 for TRPV1 K822R+SUMO1+Ubc9 (pink). k–n Representative traces of currents evoked by a family of voltage steps from −100 to +200 mV with 20 mV increments as shown in inset in k for CHO-K1 cells that expressed Flag-TRPV1 (k), Flag-TRPV1+GFP-SUMO1+Ubc9 (l), Flag-TRPV1-K822R (m), and Flag-TRPV1-K822R+GFP-SUMO1+Ubc9 (n). Holding potential was −60 mV. o Conductance–voltage (G–V) relationships derived from the experiments shown in k–m fitted with the Boltzmann function, which yielded the following results: for TRPV1 (black), V_1/2 = 105.8 ± 2.2 mV, and κ = 35.5 ± 1.9 (n = 7); TRPV1+SUMO1+Ubc9 (blue), V_1/2 = 101.5 ± 4.2 mV, and κ = 40.2 ± 3.7 (n = 7); TRPV1-K822R (olive), V_1/2 = 101.3 ± 3.7 mV, and κ = 38.9 ± 1.7 (n = 7); and TRPV1-K822R+SUMO1+Ubc9 (pink), V_1/2 = 104.8 ± 1.5 mV, and κ = 35.1 ± 1.3 (n = 5). Data points are means ± s.e.m